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Chemical Synthesis/Semisynthesis of Post-Translational Modified Tau Protein

Yehjun Lim , Yanmei Li

Progress in Chemistry ›› 2022, Vol. 34 ›› Issue (8) : 1645 -1660.

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Progress in Chemistry ›› 2022, Vol. 34 ›› Issue (8) : 1645-1660 DOI: 10.7536/PC211012
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Chemical Synthesis/Semisynthesis of Post-Translational Modified Tau Protein
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Abstract

Tau protein is a microtubule-associated protein that contains six isoforms and consists 352~441 amino acids. Misfolding and aggregation of Tau protein are closely related to Tauopathies, including Alzheimer’s disease (AD). In the brain of AD patients, it is indicated that post-translational modification plays a key factor in the pathogenesis of AD. This paper reviews the post-translational modifications of Tau protein and the progress in chemical synthetic/semisynthetic preparation of uniform Tau protein with specific site modification. Reviewing the research of post-translational modification of Tau protein, clarifies the regulatory mechanism of post-translational modification, which will help us to understand the physiological and pathological effects of post-translational modification on Tau protein and further develop potential disease treatment methods.

Contents

1 Introduction

2 The structure and function of Tau protein

3 Post-translational modification of Tau protein

3.1 Phosphorylation

3.2 Glycosylation and glycation

3.3 Acetylation

3.4 Ubiquitination

3.5 Nitration

3.6 Other post-translational modifications

4 Preparation of post-translationally modified Tau protein by chemical synthetic/semisynthetic

4.1 Preparation of post-translationally modified Tau protein microtubule binding domain by chemical synthesis

4.2 Preparation of post-translationally modified full length Tau protein by chemical synthetic/semisynthetic

5 Conclusion and outlook

Graphical abstract

Keywords

Tau protein / Tauopathies / post-translational modification / protein synthesis / protein semisynthetic

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Yehjun Lim, Yanmei Li. Chemical Synthesis/Semisynthesis of Post-Translational Modified Tau Protein. Progress in Chemistry, 2022, 34(8): 1645-1660 DOI:10.7536/PC211012

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1 Introduction

At present, the number of patients with neurodegenerative diseases worldwide has begun to exceed the number of patients with cancer, and will continue to increase rapidly in the coming decades[1,2]. Neurodegenerative diseases can lead to cognitive decline, memory loss, limb movement disorders and so on, which have a huge impact on the health and quality of life of patients. Tauopathies are a group of neurodegenerative diseases, including Alzheimer's disease (AD), frontotemporal dementia with Parkinson's disease (FTDP-17), Pick's disease (PiD), and progressive supranuclear palsy (PSP)[3]. A common feature of these diseases is the deposition of Tau protein in the brain of patients, such as neurofibrillary tangles formed by Tau protein, which is one of the main pathological signs of AD[4,5]. Studies have found that tauopathies are heterogeneous neurodegenerative diseases, that is, although tauopathies are caused by the deposition of Tau protein, they have different clinical symptoms, such as memory decline in AD patients and memory decline rarely in PiD patients[3,6].
Tau is a microtubule-associated protein that is mainly expressed in the central nervous system[7]. Tau protein is highly abundant in neuronal cell axons, whereas it is almost undetectable in astrocytes and oligodendrocytes[8]. Under physiological conditions, the main function of Tau protein is to promote the assembly of tubulin and maintain the stability of tubulin[8]. Under pathological conditions, Tau protein will aggregate to form insoluble filamentous aggregates and assemble into fibrous aggregates, resulting in tauopathy[9,10].
Tau protein has abundant post-translational modification sites, which is an important way to regulate the structure and function of Tau protein, and abnormal post-translational modification is closely related to tauopathies[11⇓⇓~14]. A large number of studies have shown that abnormal phosphorylation plays an important role in the pathogenesis of tauopathies, but not all phosphorylation sites cause Tau protein aggregation[15⇓~17][18]. In addition, other post-translational modifications (such as acetylation and nitration) may also lead to Tau protein aggregation[19⇓⇓~22]. The heterogeneity of tauopathies may be closely related to differences in post-translational modifications.
By studying the post-translational modification of Tau protein, we can analyze the differences between Tau protein aggregates, understand the molecular mechanism of the occurrence and development of Tau proteinopathy, and provide better prevention and treatment methods[23,24]. Therefore, a series of methods have been developed to prepare Tau proteins with different post-translational modifications. In this review, we will review the recent methods of preparing post-translationally modified Tau proteins, review the latest research results of post-translationally modified Tau proteins, and elaborate the regulatory mechanism of post-translational modifications on Tau proteins.

2 Structure and Function of Tau Protein

Tau protein is encoded by a single gene located on chromosome 17q21.31. Due to the alternative splicing of exons, six Tau protein isoforms with a length of 352-441 amino acids can be produced in the adult brain (fig. 1A)[25]. These subtypes are distinguished by fragments containing 0 (0 N), 29 (1 N), or 58 (2 N) amino acids at the nitrogen terminus; The carbon terminus contains three (3R) or four (4R) tandem repeats of 31-32 amino acids, resulting in six Tau protein isoforms: 0 N3R, 1 N3R, 2 N3r, 0 N4R, 1 N4r and 2 N4r[26]. In the adult brain, the ratio of 3R-Tau to 4R-Tau is approximately 1:1, while 0 N-Tau, 1 N-Tau, and 2 N-Tau account for 37%, 54%, and 9%, respectively[27].
Under physiological conditions, Tau protein has a low content of α-helix and β-sheet conformations, and mainly exists in the form of natural unfolding or inherent disorder, which is a soluble protein with strong hydrophilicity[28,29]. Tau protein has three major domains: an acidic nitrogen terminus, a proline-rich region, and a basic carbon terminus. The carbon terminus of Tau protein binds to microtubules and is called the "Assembly domain"[30]. Studies have shown that 4R-Tau has a stronger binding force with microtubules than 3R-Tau, which can promote microtubule assembly more effectively[27]. The middle region of Tau protein is rich in proline, which is the binding site of many proline-directed kinase targets and proteins with Src homology domain[31]. The nitrogen terminus of Tau protein does not bind to microtubules and is called the "Projection domain". The nitrogen terminus of Tau protrudes from the microtubule surface to interact with other cytoskeletal, mitochondrial, or neuronal plasma membranes[32].
Tau protein is a multifunctional protein, which can bind to many small molecules and biomacromolecules, including signaling molecules, cytoskeleton and cellular lipids[33⇓⇓~36]. The primary function of Tau protein is to interact with tubulin, including binding to tubulin and promoting and stabilizing the formation of microtubule assemblies[37]. In addition, Tau protein can regulate cell shape and motility, and regulate the activity of actin binding protein[38].

3 Post-translational modification of Tau protein

The post-translational modification of Tau protein is tightly regulated to regulate the structure and function of Tau protein. Post-translational modifications on Tau protein include phosphorylation, glycosylation, acetylation, ubiquitination, etc[11,39⇓~41]. Under pathological conditions, the abnormal post-translational modification of Tau protein can affect the physiological function of Tau protein, for example, phosphorylation can change the conformation of Tau protein and promote the shedding of other microtubules[42][43,44]. The formation of neurofibrillary tangles in AD is closely related to the abnormal post-translational modification of Tau protein, so a better understanding of the post-translational modification of Tau protein will help to understand the occurrence and development of Tau-related diseases, and lay the foundation for the study of related diagnosis and treatment strategies.

3.1 Phosphorylation

Phosphorylation is the most common post-translational modification of proteins. Phosphorylation can occur on serine (Ser), threonine (Thr), and tyrosine (Tyr) residues in proteins. It has been shown that the longest Tau isoform in adult brain (2N4R) contains a total of 85 potential phosphorylation sites (Fig. 1B), of which more than 40 phosphorylation sites have been identified in AD brains[12].
Tau protein is often abnormally hyperphosphorylated in diseases such as AD. Kinases involved in the regulation of Tau phosphorylation are divided into three main groups: proline-directed protein kinase (PDPK), non-PDPK protein kinase, and tyrosine protein kinase (TPK).

3.1.1 GSK-3

Glycogen synthase kinase-3 (GSK-3) belongs to PDPK and is a serine/threonine-specific kinase. To date, 21 serine and 10 threonine sites that can be phosphorylated by GSK-3 have been identified in the Tau protein (Table 1), and at least 25 of these sites have been determined to be associated with AD[39,45]. Analysis of samples from AD patients showed that GSK-3 levels were elevated in the samples, and the activity of GSK-3 was positively correlated with the increase of neurofibrillary tangles[70]. In addition to this, GSK-3 and neurotangles were found to colocalize in immunohistochemical studies[15,71].
Yang's team used GSK-3 to phosphorylate Ser235 and Ser404 of Tau protein, and verified that Ser235 and Ser404 were abnormal phosphorylation sites that induced the formation of helical filaments[72]. Moreno's group used heparin to enhance the activity of GSK-3 to prepare Ser262-phosphorylated Tau protein, and found that the binding force between GSK-3-phosphorylated Tau protein and microtubules was reduced[73]. Miller's team found that Tau protein phosphorylation by GSK-3 reduced its ability to reorganize the cellular microtubule network, while Tau protein mutant phosphorylated by GSK-3 (Ser199, Ser202, Ser235, Ser396 and Ser404 were mutated to Ala to prevent these sites from being phosphorylated by GSK-3) maintained its ability to reorganize the cellular microtubule network, indicating that the ability of Tau protein to regulate the formation of microtubules into microtubules was due to Ser199, Ser202, Ser235[74]. To further investigate how phosphorylation at different sites affects Tau regulation of microtubule assembly, Utton's group designed three Tau mutants (Ser199 and Ser202 to Ala, Ser235 to Ala, Ser396 and Ser404 to Ala)[75]. The results showed that phosphorylation of Ser199, Ser202 and Ser235 did not affect the ability of Tau protein to regulate microtubule assembly. Phosphorylation of Ser396 and Ser404 inhibited the ability of Tau protein to promote microtubule assembly. Johnson's group designed three Tau protein mutants (Thr231 to Ala, Thr235 to Ala, and both Thr231 and Thr235 to Ala) to study the role of GSK-3 in phosphorylating Tau proteins Thr231 and Thr235[76]. It was found that phosphorylation of Thr231 reduced the binding of Tau to microtubules, while phosphorylation of Thr235 did not affect the binding of Tau to microtubules. In addition, phosphorylation of Thr231 also attenuates the interaction between Tau and the cytoskeleton.

3.1.2 CDK5

Cyclin-dependent kinase 5 (CDK5) is a serine/threonine kinase belonging to PDPK. CDK5 can phosphorylate pathologically relevant six serine and five threonine sites on Tau protein (Table 1)[52]. The activation of CDK5 requires the binding of cofactor p35 or P25 to its subunits. The holoenzyme composed of P25 and CDK5 promotes the phosphorylation of Tau protein and reduces the binding ability of Tau protein to microtubules[77]. Knockout of CDK5 in a triple transgenic mouse model of AD reduces the number of neurofibrillary tangles in hippocampal tissue[78].
Cai Lihui's group and Kerokoski's group found that P25/p35 could regulate the activity of Tau protein phosphorylated by CDK5[77][79]. Under physiological conditions, p35 can be rapidly degraded to down-regulate the phosphorylation of Tau protein by CDK5. In pathological conditions, the stability of P25 leads to its accumulation in the brain, which promotes CDK5 to hyperphosphorylate Tau protein, and the binding force between phosphorylated Tau protein and microtubules becomes weak, leading to the occurrence of diseases[77,79].

3.1.3 JNK

C-Jun N-terminal kinase (JNK), also a PDPK, is a member of the mitogen-activated protein kinase (MAPKs) family. JNK was shown to phosphorylate 10 serine and 6 threonine sites of Tau protein (Table 1)[45]. Immunohistochemical analysis showed that JNK was colocalized with Tau protein aggregates in AD. Elevated levels of JNK were also found in the brains of AD patients, and activated JNK (p-JNK) was colocalized with Tau in neurons[80].

3.1.4 CK1

Casein kinase 1 (CK1) is a serine/threonine kinase that belongs to the non-PDPK class. Twenty serines and three threonines on Tau protein can be phosphorylated by CK1 (see Table 1)[46]. In vitro, CK1 was found to regulate the binding of Tau to microtubules by phosphorylating Ser202/Thr205 and Ser396/Ser404 of Tau. In addition, it was found that CK1 and neurofibrillary tangles were colocalized in the brain of AD patients[81]. Kuret's team found that CK1 phosphorylated Ser202, Thr205, Ser396 and Ser404 of Tau protein, which disrupted the binding between Tau protein and microtubules[81].

3.1.5 PKA

Protein kinase A (PKA), also known as cyclic AMP-dependent kinase (cAMP-PK), belongs to non-PDPK. PKA is involved in the phosphorylation of 16 serines and 4 threonines of Tau (see Table 1), and Tau phosphorylated by PKA triggers further phosphorylation of Tau by GSK-3 and CDK5[57,58]. Johnson's team found that phosphorylation of Tau protein by PKA can reduce the binding of Tau protein to microtubules and improve the ability of Tau protein to resist proteolytic degradation, resulting in the accumulation of Tauprotein[82⇓~84]. Later, they found that PKA regulates the ability of Tau to bind to microtubules and resist degradation by phosphorylating Ser262 and Ser356 of Tau[85]. Mandelkow's team found that PKA phosphorylated Ser214 of Tau protein could inhibit the formation of helical filaments of Tau protein[65].

3.1.6 DYRK

Dual specificity tyrosine phosphorylation-regulated kinase (DYRK) is found in neurons of the cortex and hippocampus of AD patients. DYRK can phosphorylate six serine and five threonine sites on Tau protein (Table 1). Phosphorylation of DYRK can cause GSK-3 to further phosphorylate Thr181, Ser199, Ser202, Thr205 and Ser208 of Tau protein, but DYRK is not involved in the regulation of CDK5[61⇓~63]. Up-regulating the activity of DYRK will lead to abnormal phosphorylation of Tau protein, which was confirmed in model mouse experiments. DYRK-induced phosphorylation inhibits the activity of Tau protein in regulating microtubule assembly and promotes Tau protein aggregation[16]. Song's team found that DYRK can phosphorylate Ser202 and Ser404 of Tau protein, thus strongly inhibiting the ability of Tau protein to promote microtubule assembly[61].

3.1.7 MARKs

Microtubule binding force-regulating kinases (MARKs) belong to non-PDPK. MARKs have been found to be elevated in the brain of AD patients and colocalize with neurofibrillary tangles[64]. MARKs phosphorylate mainly 4 serines located in the repeat domain in Tau protein (see Table 1)[86,87]. Mandelkow's team found that MARK can phosphorylate four sites in the tandem repeat sequence of Tau protein, namely Ser262, Ser293, Ser324 and Ser356[65]. They found that phosphorylation of Tau protein at Ser262 leads to the shedding of Tau protein from microtubules, which affects the stability of microtubules[65,88].

3.1.8 AMPK

Non-PDPK AMPK is a heterotrimeric serine/threonine kinase. In vitro studies have shown that AMPK phosphorylates 15 serine and 8 threonine sites of Tau protein (Table 1)[66]. Dickson's team found that AMPK phosphorylated Tau protein Thr231 and Ser396/Ser404 could promote Tau protein to form neurofibrillary tangles[66].

3.1.9 SFK

Src protein kinase family (SFK) belongs to TPK, and its members include Fyn protein kinase and Lck protein kinase. Masliah's team found that Fyn protein levels in the brain of AD patients were elevated and associated with cognitive impairment[89].
In 1993, Wood's team found that the brain of AD patients had higher levels of Fyn compared with healthy brains[90]. Gloria's team found that Fyn can phosphorylate Tyr18 of Tau protein and affect the binding between Tau protein and microtubules[91]. Reynolds' group identified that Fyn protein kinase could phosphorylate Tyr18 and Tyr197 of Tau protein, and Lck could phosphorylate Tyr18, Tyr197, Tyr310 and Tyr394[68][68].

3.1.10 Abl

The Abl protein kinase family (Abl) is a receptor-independent TPK. The c-Abl protein kinase in the Abl protein kinase family can phosphorylate Tyr197, Tyr310, and Tyr394 of Tau protein[69]. Immunohistochemical studies by Matthew's group have shown that c-Abl phosphorylates Tyr394 of Tau protein, which is related to the formation of nerve plaques and neurofibrillary tangles in AD patients[17]. In model mice, overexpression of c-Abl leads to phosphorylation of Tau protein and neurodegeneration. Lashuel's team phosphorylated Tyr310 of Tau protein by c-Abl, and found that phosphorylation of this site could inhibit the aggregation of Tau protein and weaken its binding to microtubules and lipids[92].

3.2 Glycosylation and glycation

Glycosylation is the covalent binding of proteins and oligosaccharides by glycosyltransferases. Glycosylation is another important form of post-translational modification for Tau protein. Glycosylation can be divided into O-glycosylation and N-glycosylation, in which the amino group of the side chain of asparagine (Asn) is the most common N-glycosylation reaction site, while the hydroxyl group of the side chain of serine/threonine is the most common O-glycosylation reaction site.
Tau protein contains 15 O-glycosylation sites, including Thr123, Thr181, Ser199, Ser202, Thr205, Ser208, Thr212, Ser214, Thr217, Ser238, Ser262, Ser256, Ser400, Ser404, and Ser422[93⇓~95]. O-glycosylation modification is associated with N-acetyl-D-glucosamine polypeptide transferase (OGT). OGT catalyzes the reaction of β-N-acetylglucosamine (GlcNAc) with the side chain hydroxyl group on serine/threonine to form O-glycosylation modification. It has been shown that there is a reciprocal regulatory relationship between phosphorylation and O-glycosylation on Tau protein[96].
Glycation is a nonenzymatic glycosylation modification of proteins frequently observed during tissue aging (Scheme 1), and is the product of a reaction between a monosaccharide, such as glucose or fructose, and a lysine (Lys) side chain amino group on the protein[97]. A total of 12 glycosylation sites were found on Tau protein, including Lys87, Lys132, Lys150, Lys163, Lys 174, Lys 225, Ly[98⇓~100]. Significant amounts of AGEs were found in neurofibrillary tangles from AD patient samples[101,102]. Glycation was found to promote Tau protein aggregation and stabilize Tau protein aggregates, but not to cause Tau protein spontaneous fibrillation or increase the nucleation rate[99]. Glycation may inhibit the degradation of Tau protein, which leads to the accumulation of Tau protein and the death of neuronal cells[103,104].

3.3 Acetylation

Acetylation occurs mainly on the N-terminal amino group or lysine side chain amino group of the protein and is catalyzed by histone acetyltransferase (HAT) or lysine acetyltransferase (Scheme 2)[105]. In 2010, Gan group identified that Tau protein contains 23 potential acetylation sites in vitro using acetyltransferase P300 or pCAF, among which Lys174, Lys274 and Lys281 have special pathological significance[106]. Studies have found increased levels of Tau protein acetylation in the brains of AD patients. Further mechanistic studies have shown that acetylation of Tau protein can inhibit its degradation and binding to microtubules, and promote the aggregation of Tau protein[106][19][19,20].

3.4 Ubiquitination

Ubiquitination is the covalent binding of a ubiquitin protein to a target protein, and the protein modified by ubiquitination is degraded by the ubiquitin-proteasome system (UPS) in the cytoplasm. Mass spectrometry and immunohistochemistry have found that there are single/polyubiquitin chains (Lys6-, Lys11-, Lys48- and Lys63-) between ubiquitin and Tau protein[107,108]. At present, Lys254, Lys257, Lys311 and Lys317 have been identified as monoubiquitination sites on Tau protein, while Lys254, Lys 311 and Lys 353 are polyubiquitin chain modification sites[107,109]. It has been reported that a variety of E3 ligases can ubiquitinate Tau protein, including heat shock protein 70 carbon terminal interacting protein (CHIP) and tumor necrosis factor receptor-associated factor 6 (TRAF6)[110⇓⇓~113]. The ubiquitination of Tau proteins Lys254, Lys311 and Lys353 can weaken the binding force of Tau proteins to microtubules, and Tau proteins can be detached from microtubules and enter the degradation process. Proteasome was found to co-precipitate with Tau protein in neurofibrillary tangles isolated from the brains of AD patients. Studies have shown that proteasome processing efficiency and degradation activity are significantly impaired in AD patients, so that Tau protein can not be effectively degraded[114,115].

3.5 Nitration

3-Nitrotyrosine is considered to be a biomarker of endogenous nitrosylation and nitrosylation of proteins and is one of the causes of neuronal injury and cell degeneration[116][117]. The formation of 3-nitrotyrosine may be related to the increase of intracellular oxidative stress, but the mechanism has not yet been clarified[118]. Only four tyrosines (Tyr18, Tyr29, Tyr197 and Tyr394) of the five tyrosines in Tau protein can be modified by nitration, while the nitration of Tyr310 has not been reported[119]. Nitrosylation of Tyr18 and Tyr197 is found in both healthy and AD brains and may not be of pathological significance[120,121]; Nitrosylation of Tyr29 is found only in the brains of AD patients[22]. In vitro experiments showed that nitrosylation could weaken the interaction between Tau protein and microtubules and cause Tau protein polymerization[22].

3.6 Other post-translational modification

In addition to the above post-translational modifications, Tau protein also has post-translational modifications such as ubiquitination-like (SUMO), oxidation, peptide proline isomerization, truncation, and polyamination[122,123][124][125][126][127].
Tau can be sumoylated on K340 in vitro[122,123]. SUMOylation of Tau protein can induce hyperphosphorylation of multiple sites in Tau protein and inhibit the degradation of Tau protein[128]. In addition, SUMOylated Tau can be detected in AD brains in coexistence with hyperphosphorylated Tau[123].
Cys322 of Tau protein can be oxidized, which leads to the assembly of neurofibrillary tangles[124,129]. At present, there are few studies on the oxidative modification of Tau protein, and the relationship between Cys322 oxidation of Tau protein and disease is still unclear.
The pSer/Thr-Pro sequence in the protein has both cis and trans conformations, and peptidyl isomerization can occur under the action of peptidyl isomerase (Pin1). There are also two Pin1 recognition sites in Tau, pThr212-Pro213 and pThr231-Pro232[125]. It is believed that Tau protein in the trans conformation is more easily dephosphorylated, thus restoring the ability of Tau protein to bind to microtubules[125,130].
Transglutaminase can catalyze the reaction of lysine and glutamine (Gln) to form an isopeptide bond, which is called polyamination (Scheme 3) and can lead to protein cross-linking[131]. Ten donor sites (major donor sites are Lys163 and Lys240) and eight acceptor sites (major acceptor sites are Gln351 and Gln424) were identified on Tau protein[127]. Studies have shown that polyamination of Tau protein may be involved in the formation of neurofibrillary tangles[132,133].

4 Chemical full/semisynthetic preparation of post-translationally modified Tau protein

It has been described above that Tau protein has multiple types of post-translational modifications and that the same post-translational modification has multiple modification sites. In order to study the effects of post-translational modifications on the structure and function of Tau protein, it is necessary to obtain proteins with clear modification sites and uniform structure. It is difficult to extract samples directly from the patient's brain and perform a series of purifications to obtain homogeneous and structurally defined proteins with specific post-translational modifications. At the same time, after many rounds of purification, the sample size is very small, and it is difficult to carry out various characterizations to fully understand the effects of post-translational modifications on proteins. In order to study the effects of post-translational modifications on proteins more deeply, it is necessary to develop preparation methods that can obtain large amounts of target proteins. At present, there are two main methods to prepare post-translationally modified proteins in vitro: biological enzymatic method and chemical full/semi-synthesis.
Enzymatic methods mainly use E. coli to express recombinant proteins to obtain a large number of wild-type proteins, and then modify them by related proteases to obtain proteins with post-translational modifications. Enzymatic method has the advantages of high yield, fast speed and high molecular weight protein production. Because of the broad spectrum of proteases, proteases may modify multiple sites at the same time, so it is impossible to directly obtain the target protein modified by a single site through biological enzymatic methods. In order to obtain a single-site modified target protein, other potential post-translational modification sites need to be mutated to other amino acids (such as alanine) and then post-translationally modified by proteases, but the mutation may affect the overall structure of the protein.
The type or site of post-translational modification has different effects on the structure and properties of amyloid proteins[18,134⇓⇓⇓~138]. In order to clarify the influence of specific modifications at a certain site on the properties of Tau protein, it is necessary to prepare Tau protein with clear modification types and sites. Compared with biological enzymatic methods, chemical synthesis is easier to introduce modifications at specific sites, which is conducive to better study how modifications at specific sites regulate the properties of proteins.
The natural chemical ligation (NCL) method developed by Kent's group and the expressed protein ligation (EPL) method developed by Muir's group can realize chemical total synthesis/semi-synthesis of proteins. The target protein can be obtained by introducing structural units with specific modifications (such as phosphorylation, glycosylation, etc.) During the synthesis process through the solid-phase peptide synthesis (SPPS), and then coupling the polypeptide fragments through the natural chemical ligation method and the expressed protein ligation method (Figure 4). At present, many proteins with specific post-translational modifications have been synthesized by natural chemical ligation and expression ligation for structural and biological studies[139⇓⇓~142].

4.1 Preparation of post-translationally modified microtubule binding domain of Tau protein by chemical synthesis

4.1.1 Preparation of phosphorylated microtubule binding domain of Tau protein by chemical synthesis

The microtubule-binding domain of Tau consists of 3 or 4 tandem repeats (Tau3RD or Tau4RD). Lashuel's group used the three-fragment method to synthesize singly phosphorylated (pS356 Tau4RD), doubly phosphorylated (pS262 & pS356 Tau4RD), and triply phosphorylated Tau4RD (pS258 & pS262 & pS356 Tau4RD) (Scheme 5)[18]. It was found that the aggregation ability of Tau4RD became weaker as the amount of phosphorylation increased[18]. In addition, phosphorylation of serine 262 was found to significantly disrupt the binding ability of Tau protein to microtubules[18].

4.1.2 Preparation of ubiquitinated Tau protein microtubule binding domain by chemical synthesis

The Onofrio group used Tau4RD as a model to study ubiquitinated Tau protein[143,144]. Their strategy was to link ubiquitin to Tau protein through disulfide bonds: first, two cysteines on Tau4RD were mutated to alanine (C291A and C322A), and then the lysines on Tau4RD were mutated to cysteines (K254C/K311C/K353C), respectively, and then the three mutated cysteines were linked to ubiquitin to obtain three monoubiquitin-modified Tau4RDs (Ub254 Tau4RD, Ub311 Tau4RD, and Ub 35. It was found that Ub311 Tau4RD could change the conformation of Tau protein and inhibit the formation of fibers. Ub254 Tau4RD and Ub353 Tau4RD can form fibers, but the aggregation process is significantly slowed down compared with the wild type Tau4RD protein. To further examine the effect of diubiquitination on Tau protein, they constructed diubiquitin molecules linked in both Lys48- and Lys63-ways (48-diUb and 63-diUb). Next, 48-diUb353 Tau4RD and 63-diUb353 Tau4RD were constructed by the same disulfide strategy (fig. 6B). It was found that diubiquitinated Tau protein was more prone to form oligomers than fibers. In addition, diubiquitination also slowed down the aggregation rate, but did not inhibit it.

4.2 Preparation of post-translationally modified full-length Tau protein by chemical full/semisynthesis

4.2.1 Preparation of phosphorylated and acetylated Tau protein by chemical full/semisynthesis

In 2012, Hackenberger's group used chemical semisynthesis to synthesize full-length Tau protein[145]. They obtained recombinant protein thioester (Tau 1-389) by protein expression, used phosphorylated serine to synthesize phosphorylated Tau 390-441 fragment, and connected the two fragments by NCL to obtain monophosphorylated Tau protein at position 404 (pS404 Tau) (Scheme 7). To better purify the synthetic protein, they introduced biotin at Lys438 and purified it by affinity column chromatography. Studies have shown that the introduction of A390C in the synthesis process does not affect the properties of the protein. In addition, the experimental results showed that serine 404 phosphorylation did not alter the ability of Tau protein to promote microtubule polymerization.
In 2018, Lashuel's group used a five-fragment method to synthesize Tau protein with post-translational modifications, which were 2-245, 246-290, 291-321, 322-389 and 390-441, respectively[21]. In this strategy, each fragment from 246 to 441 was obtained by chemical synthesis, covering the microtubule-binding region and the carbon terminus of Tau protein, so this strategy can introduce many disease-related post-translational modifications at specific locations. Based on this strategy, Lashuel's group synthesized Tau protein monophosphorylated at tyrosine 310 (pY310 Tau), Tau protein bisphosphorylated at serine 396 and serine 404 (pS396 & pS404 Tau) (Figure 8), and Tau protein acetylated at lysine 280 (AcK280 Tau) (Figure 9)[21]. It was found that the aggregation rate of AcK280 Tau protein was much higher than that of wild type Tau. Wild-type Tau protein mainly aggregated to form insoluble fibrils, while AcK280 Tau protein formed soluble aggregates. Acetylation of lysine at position 280 promotes Tau protein oligomerization to form short fibers. In addition, AcK280 Tau protein was found to inhibit the assembly of tubulin to form microtubules.
In order to obtain modified Tau protein more efficiently for the study of post-translational modification of Tau protein microtubule binding region, Lashuel's group expressed Tau 1-290 and Tau 322-441 by recombinant protein, chemically semi-synthesized Tau 291-321, and then connected the three fragments by NCL[146]. Using this strategy, they synthesized Tau proteins phosphorylated at serine 293 (pS293 Tau) and serine 305 (pS305 Tau) (Scheme 10). pS293 Tau was found to aggregate slightly faster than WT Tau protein, whereas pS305 Tau aggregated more slowly.

4.2.2 Preparation of glycosylated Tau protein by chemical semisynthesis

Hackenberger's group synthesized Tau protein with glycosylation modification in 2016[147]. They expressed one fragment (Tau 1 ~ 389) by recombinant protein and chemically synthesized two fragments (gS400 Tau 390 ~ 410 and Tau 411 ~ 441). Next, the three fragments were ligated by NCL. Through this strategy, they obtained a Tau protein modified with serine O-GlcNac at position 400 (Scheme 11).

4.2.3 Preparation of nitrated full-length Tau protein by in vitro modification

In order to obtain Tau protein modified by nitration at specific sites to study its properties, Reynolds's group prepared Tau protein mutants that only retained the nitrosomimetic tyrosine and mutated the other tyrosines to alanine, and then used free radical peroxynitrite to nitrate the retained tyrosine.Tau proteins with tyrosine nitration at position 18 (nTyr18 Tau), tyrosine nitration at position 29 (nTyr29 Tau), tyrosine nitro-ation at position 197 (nTyr197 Tau) and tyrosine nitro-ation at position 394 (nTyr 394 Tau) were obtained, respectively[119]. It was found that Tyr18 and Tyr29 located at the nitrogen terminus were more likely to undergo nitration than Tyr197 and Tyr394. In addition, the average length of nTyr29 Tau and nTyr197 Tau aggregation fibers was longer than that of wild type Tau protein, while the average length of nTyr18 Tau and nTyr394 Tau aggregation fibers was shorter than that of wild type Tau protein[119]. Thus, the location of nitration has a strong influence on the aggregation properties. In addition, the nitrosylation of Tyr18 and Tyr394 increases the critical concentration of protein aggregation, which promotes the aggregation equilibrium to the monomeric state[119]. In 2006, Reynolds' group further found that the nitration of Tyr18, Tyr29, Tyr197 and Tyr394 of Tau protein may lead to the weakening of the binding ability of Tau protein with microtubules, resulting in the lack of stability of microtubules and Tau protein assemblies, and ultimately affecting the function of the assemblies[148].

5 Summary and Prospect

Some progress has been made in understanding the relationship between Tau protein and tauopathies. Abnormal post-translational modification of Tau protein is the main cause of Tau protein forming neurofibrillary tangles. However, it is still difficult to clearly understand the molecular mechanism of the effects of specific post-translational modifications on Tau protein at some sites. Early studies mainly focused on the modification of Tau protein by proteases, but some proteases did not have site specificity for the modification of Tau protein, resulting in the formation of a mixture of different sites, so that the role of a single site could not be known. Post-translational modifications are mimicked by using mutants, such as serine to aspartic acid or glutamic acid to mimic phosphorylated Tau protein, but the mutants do not fully exhibit the true properties of the native modification. Chemical full/semisynthesis provides powerful strategies to generate site-specifically modified Tau proteins, including phosphorylation, glycosylation, acetylation, ubiquitination, etc.
Many types of post-translational modifications can occur at many sites of Tau protein, and the competition and regulatory relationships between the different modifications at these sites are still unclear or controversial and require further investigation[96,149,150]. For example, Miller's group phosphorylated Tau protein by GSK-3, and found that phosphorylation of Tau protein Ser396 and Ser404 may inhibit the ability of Tau protein to promote microtubule assembly[75]. Because GSK-3 can phosphorylate many sites, it is not clear whether Ser396 or Ser404 phosphorylation is the main cause of Tau protein inhibition. Hackenberger's group prepared pS404 Tau by chemical synthesis, which clearly showed that phosphorylation at position 404 did not affect the function of Tau protein[145]. Therefore, the effect of post-translational modification on proteins can be more clearly understood by preparing specific modified proteins through chemical synthesis[151]. These results provide important evidence for revealing the endogenous function of Tau protein and the pathogenesis of tauopathy, and provide a theoretical basis for the treatment of tauopathy.
Although chemical full/half synthesis provides a powerful tool for the study of Tau protein, there are still some problems to be solved. First, chemical full/semisynthesis usually has low yields due to technical limitations. Therefore, it is necessary to develop new synthesis and purification methods to produce post-translationally modified Tau protein more rapidly and conveniently. Secondly, different post-translational modifications have different effects on the properties of Tau protein (including aggregation and toxicity), but the types of modified Tau protein synthesized are limited, and there are few in-depth studies on it. In a word, chemical full/semisynthesis can help people understand Tau protein more systematically and deeply, provide insights into the post-translational modification of Tau protein, and lay the foundation for the development of new strategies for the treatment and prevention of Tauopathies.

References

Publishing order | Descend order by publishing year | Descend order by cited within
[1]

Sierksma A, Escott-Price V, de Strooper B. Science, 2020, 370(6512): 61.
[2]

Li G, Li Y M. Progress in Chemistry, 2020, 32 (1): 14.
[3]

Kovacs G G. Neuropathology, 2018, 145: 355.
[4]

Goedert M, Spillantini M G. Science, 2006, 314(5800): 777.
[5]

Lee V M Y, Balin B J, Otvos L Jr, Trojanowski J Q. Science, 1991, 251(4994): 675.
[6]

Lebouvier T, Pasquier F, BuÉe L. Curr. Opin. Neurol., 2017, 30(6): 589.
[7]

Goedert M, Spillantini M G, Jakes R, Rutherford D, Crowther R A. Neuron, 1989, 3(4): 519.
[8]

Avila J, Gomez de Barreda E, Pallas-Bazarra N, Hernandez F. Aging and Disease, 2013, 4 (1): 23.
[9]

Bancher C, Brunner C, Lassmann H, Budka H, Jellinger K, Wiche G, Seitelberger F, Grundke-Iqbal I, Iqbal K, Wisniewski H M. Brain Res., 1989, 477(1/2): 90.
[10]

Alonso A D C, Zaidi T, Novak M, Grundke-Iqbal I, Iqbal K. PNAS, 2001, 98(12): 6923.
[11]

Martin L, Latypova X, Terro F. Neurochem. Int., 2011, 58(4): 458.
[12]

Wang J Z, Xia Y Y, Grundke-Iqbal I, Iqbal K. J. Alzheimer’s Dis., 2013, 33(s1): S123.
[13]

Park S, Lee J H, Jeon J H, Lee M J. BMB Rep., 2018, 51(6): 265.
[14]

Chen Z Z, Tan B, Li Y M, Zhao Y F, Tong Y F, Wang J F. J. Org. Chem., 2003, 68(10): 4052.
[15]

Yamaguchi H, Ishiguro K, Uchida T, Takashima A, Lemere C A, Imahori K. Acta Neuropathol., 1996, 92(3): 232.
[16]

Ferrer I, Barrachina M, Puig B, Martí E, Avila J, Dierssen M. Neurobiol. Dis., 2005, 20(2): 392.
[17]

Tremblay M A, Acker C M, Davies P. J. Alzheimer’s Dis., 2010, 19(2): 721.
[18]

Haj-Yahya M, Gopinath P, Rajasekhar K, Mirbaha H, Diamond M I, Lashuel H A. Angew. Chem. Int. Ed., 2020, 59(10): 4059.
[19]

Cohen T J, Guo J L, Hurtado D E, Kwong L K, Mills I P, Trojanowski J Q, Lee V M Y. Nat. Commun., 2011, 2: 252.
[20]

Min S W, Chen X, Tracy T E, Li Y Q, Zhou Y G, Wang C, Shirakawa K, Minami S S, Defensor E, Mok S A, Sohn P D, Schilling B, Cong X, Ellerby L, Gibson B W, Johnson J, Krogan N, Shamloo M, Gestwicki J, Masliah E, Verdin E, Gan L. Nat. Med., 2015, 21(10): 1154.
[21]

Haj-Yahya M, Lashuel H A. J. Am. Chem. Soc., 2018, 140(21): 6611.
[22]

Reynolds M R, Reyes J F, Fu Y, Bigio E H, Guillozet-Bongaarts A L, Berry R W, Binder L I. J. Neurosci., 2006, 26(42): 10636.
[23]

Liang C Q, Li Y M. Curr. Opin. Chem. Biol., 2021, 64: 124.
[24]

Liu P, Zhang S, Chen M S, Liu Q, Wang C X, Wang C, Li Y M, Besenbacher F, Dong M D. Chem. Commun., 2012, 48(2): 191.
[25]

Goedert M, Wischik C M, Crowther R A, Walker J E, Klug A. PNAS, 1988, 85(11): 4051.
[26]

Goedert M, Spillantini M G, Potier M C, Ulrich J, Crowther R A. EMBO J., 1989, 8(2): 393.
[27]

Goedert M, Jakes R. EMBO J., 1990, 9(13): 4225.
[28]

Jeganathan S, von Bergen M, Mandelkow E M, Mandelkow E. Biochemistry, 2008, 47(40): 10526.
[29]

Mukrasch M D, Bibow S, Korukottu J, Jeganathan S, Biernat J, Griesinger C, Mandelkow E, Zweckstetter M. PLoS Biol., 2009, 7(2): e1000034.
[30]

Steiner B, Mandelkow E M, Biernat J, Gustke N, Meyer H E, Schmidt B, Mieskes G, Söling H D, Drechsel D, Kirschner M W. EMBO J., 1990, 9(11): 3539.
[31]

Mandelkow E M, Mandelkow E. Cold Spring Harb. Perspect. Med., 2012, 2(7): a006247.
[32]

Al-Bassam J, Ozer R S, Safer D, Halpain S, Milligan R A. Journal of Cell Biology, 2002, 157 (7): 1187.
[33]

BuÉe L, Bussière T, BuÉe-Scherrer V, Delacourte A, Hof P R. Brain Res. Rev., 2000, 33(1): 95.
[34]

Kosik K S. Brain Pathol., 1993, 3(1): 39.
[35]

Wang J Z, Liu F. Prog. Neurobiol., 2008, 85(2): 148.
[36]

Guo W T, Stoklund Dittlau K, van den Bosch L. Semin. Cell Dev. Biol., 2020, 99: 133.
[37]

Dolan P J, Johnson G V W. Current Opinion in Drug Discovery & Development, 2010, 13 (5): 595.
[38]

Shahani N, Brandt R. Cell. Mol. Life Sci. CMLS, 2002, 59(10): 1668.
[39]

Hanger D P, Anderton B H, Noble W. Trends Mol. Med., 2009, 15(3): 112.
[40]

Gong C X, Liu F, Grundke-Iqbal I, Iqbal K. J. Neural Transm., 2005, 112(6): 813.
[41]

Sergeant N, Bretteville A, Hamdane M, Caillet-Boudin M L, Grognet P, Bombois S, Blum D, Delacourte A, Pasquier F, Vanmechelen E, Schraen-Maschke S, BuÉe L. Expert Rev. Proteom., 2008, 5(2): 207.
[42]

Avila J. J. Cell. Mol. Med., 2008, 12(1): 258.
[43]

Fischer D, Mukrasch M D, Biernat J, Bibow S, Blackledge M, Griesinger C, Mandelkow E, Zweckstetter M. Biochemistry, 2009, 48(42): 10047.
[44]

Jho Y S, Zhulina E B, Kim M W, Pincus P A. Biophys. J., 2010, 99(8): 2387.
[45]

Reynolds C H, Betts J C, Blackstock W P, Nebreda A R, Anderton B H. J. Neurochem., 2000, 74(4): 1587.
[46]

Hanger D P, Byers H L, Wray S, Leung K Y, Saxton M J, Seereeram A, Reynolds C H, Ward M A, Anderton B H. J. Biol. Chem., 2007, 282(32): 23645.
[47]

Hanger D P, Hughes K, Woodgett J R, Brion J P, Anderton B H. Neurosci. Lett., 1992, 147(1): 58.
[48]

Mandelkow E M, Drewes G, Biernat J, Gustke N, van Lint J, Vandenheede J R, Mandelkow E. FEBS Lett., 1992, 314(3): 315.
[49]

Ishiguro K, Omori A, Takamatsu M, Sato K, Arioka M, Uchida T, Imahori K. Neurosci. Lett., 1992, 148(1/2): 202.
[50]

Yang S D, Yu J S, Shiah S G, Huang J J. J. Neurochem., 1994, 63(4): 1416.
[51]

Song J S, Yang S D. J. Protein Chem., 1995, 14(2): 95.
[52]

Paudel H K, Lew J, Ali Z, Wang J H. J. Biol. Chem., 1993, 268(31): 23512.
[53]

Lund E T, McKenna R, Evans D B, Sharma S K, Mathews W R. J. Neurochem., 2001, 76(4): 1221.
[54]

Liu F, Iqbal K, Grundke-Iqbal I, Gong C X. FEBS Lett., 2002, 530(1/3): 209.
[55]

Reynolds C H, Utton M A, Gibb G M, Yates A, Anderton B H. J. Neurochem., 1997, 68(4): 1736.
[56]

Yoshida H, Hastie C J, McLauchlan H, Cohen P, Goedert M. J. Neurochem., 2004, 90(2): 352.
[57]

Liu F, Liang Z H, Shi J H, Yin D M, El-Akkad E, Grundke-Iqbal I, Iqbal K, Gong C X. FEBS Lett., 2006, 580(26): 6269.
[58]

Liu F, Li B, Tung E J, Grundke-Iqbal I, Iqbal K, Gong C X. Eur. J. Neurosci., 2007, 26(12): 3429.
[59]

Scott C W, Spreen R C, Herman J L, Chow F P, Davison M D, Young J, Caputo C B. J. Biol. Chem., 1993, 268(2): 1166.
[60]

Hanger D P, Betts J C, Loviny T L F, Blackstock W P, Anderton B H. J. Neurochem., 1998, 71(6): 2465.
[61]

Ryoo S R, Jeong H K, Radnaabazar C, Yoo J J, Cho H J, Lee H W, Kim I S, Cheon Y H, Ahn Y S, Chung S H, Song W J. J. Biol. Chem., 2007, 282(48): 34850.
[62]

Liu F, Liang Z H, Wegiel J, Hwang Y W, Iqbal K, Grundke-Iqbal I, Ramakrishna N, Gong C X. FASEB J., 2008, 22(9): 3224.
[63]

Wegiel J, Kaczmarski W, Barua M, Kuchna I, Nowicki K, Wang K C, Wegiel J, Ma S Y, Frackowiak J, Mazur-Kolecka B, Silverman W P, Reisberg B, Monteiro I, de Leon M, Wisniewski T, Dalton A, Lai F, Hwang Y W, Adayev T, Liu F, Iqbal K, Iqbal I G, Gong C X. J. Neuropathol. Exp. Neurol., 2011, 70(1): 36.
[64]

Chin J Y, Knowles R B, Schneider A, Drewes G, Mandelkow E M, Hyman B T. J. Neuropathol. Exp. Neurol., 2000, 59(11): 966.
[65]

Schneider A, Biernat J, von Bergen M, Mandelkow E, Mandelkow E M. Biochemistry, 1999, 38(12): 3549.
[66]

Vingtdeux V, Davies P, Dickson D W, Marambaud P. Acta Neuropathol., 2011, 121(3): 337.
[67]

Thornton C, Bright N J, Sastre M, Muckett P J, Carling D. Biochem. J., 2011, 434(3): 503.
[68]

Scales T M, Derkinderen P, Leung K Y, Byers H L, Ward M A, Price C, Bird I N, Perera T, Kellie S, Williamson R, Anderton B H, Reynolds C H. Mol. Neurodegener., 2011, 6(1): 12.
[69]

Derkinderen P, Scales T M E, Hanger D P, Leung K Y, Byers H L, Ward M A, Lenz C, Price C, Bird I N, Perera T, Kellie S, Williamson R, Noble W, Van Etten R A, Leroy K, Brion J P, Reynolds C H, Anderton B H. Journal of Neuroscience, 2005, 25 (28): 6584.
[70]

Leroy K, Boutajangout A, Authelet M, Woodgett J R, Anderton B H, Brion J P. Acta Neuropathologica, 2002, 103 (2): 91.
[71]

Pei J J, Braak E, Braak H, Grundke-Iqbal I, Iqbal K, Winblad B, Cowburn R F. J. Neuropathol. Exp. Neurol., 1999, 58(9): 1010.
[72]

Yang S D, Song J S, Yu J S, Shiah S G. J. Neurochem., 1993, 61(5): 1742.
[73]

Moreno F J, Medina M, PÉrez M, Montejo de Garcini E, Avila J. FEBS Lett., 1995, 372(1): 65.
[74]

Wagner U, Utton M, Gallo J M, Miller C C. J. Cell Sci., 1996, 109(6): 1537.
[75]

Utton M A, Vandecandelaere A, Wagner U, Reynolds C H, Gibb G M, Miller C C J, Bayley P M, Anderton B H. Biochem. J., 1997, 323(3): 741.
[76]

Cho J H, Johnson G V W. J. Neurochem., 2004, 88(2): 349.
[77]

Patrick G N, Zukerberg L, Nikolic M, de la Monte S, Dikkes P, Tsai L H. Nature, 1999, 402(6762): 615.
[78]

Piedrahita D, Hernandez I, Lopez-Tobon A, Fedorov D, Obara B, Manjunath B S, Boudreau R L, Davidson B, LaFerla F, Gallego-Gomez J C, Kosik K S, Cardona-Gomez G P. J. Neurosci., 2010, 30(42): 13966.
[79]

Kerokoski P, Suuronen T, Salminen A, Soininen H, Pirttilä T. Biochem. Biophys. Res. Commun., 2001, 280(4): 998.
[80]

Zhu X W, Raina A K, Rottkamp C A, Aliev G, Perry G, Boux H, Smith M A. J. Neurochem., 2001, 76(2): 435.
[81]

Li G B, Yin H S, Kuret J. J. Biol. Chem., 2004, 279(16): 15938.
[82]

Johnson G V W. J. Neurochem., 1992, 59(6): 2056.
[83]

Litersky J M, Johnson G V. J. Biol. Chem., 1992, 267(3): 1563.
[84]

Litersky J M, Johnson G V W. J. Neurochem., 1995, 65(2): 903.
[85]

Xie H Q, Litersky J M, Hartigan J A, Jope R S, Johnson G V W. Brain Res., 1998, 798(1/2): 173.
[86]

Augustinack J C, Schneider A, Mandelkow E M, Hyman B T. Acta Neuropathol., 2002, 103(1): 26.
[87]

Mocanu M M, Nissen A, Eckermann K, Khlistunova I, Biernat J, Drexler D, Petrova O, Schonig K, Bujard H, Mandelkow E, Zhou L, Rune G, Mandelkow E M. J. Neurosci., 2008, 28(3): 737.
[88]

Drewes G, Trinczek B, Illenberger S, Biernat J, Schmittulms G, Meyer H E, Mandelkow E M, Mandelkow E. Journal of Biological Chemistry, 1995, 270 (13): 7679.
[89]

Ho G J, Hashimoto M, Adame A, Izu M, Alford M F, Thal L J, Hansen L A, Masliah E. Neurobiol. Aging, 2005, 26(5): 625.
[90]

Shirazi S K, Wood J G. NeuroReport, 1993, 4(4): 435.
[91]

Lee G. Biochim. Biophys. Acta BBA Mol. Basis Dis., 2005, 1739(2/3): 323.
[92]

Ait-Bouziad N, Chiki A, Limorenko G, Xiao S F, Eliezer D, Lashuel H A. J. Biol. Chem., 2020, 295(23): 7905.
[93]

Arnold C S, Johnson G W, Cole R N, Dong D L Y, Lee M, Hart G W. J. Biol. Chem., 1996, 271(46): 28741.
[94]

Wang J Z, Grundke-Iqbal I, Iqbal K. Nat. Med., 1996, 2(8): 871.
[95]

Yuzwa S A, Yadav A K, Skorobogatko Y, Clark T, Vosseller K, Vocadlo D J. Amino Acids, 2011, 40(3): 857.
[96]

Lefebvre T, Ferreira S, Dupont-Wallois L, Bussière T, Dupire M J, Delacourte A, Michalski J C, Caillet-Boudin M L. Biochim. Biophys. Acta BBA Gen. Subj., 2003, 1619(2): 167.
[97]

Lapolla A, Traldi P, Fedele D. Clin. Biochem., 2005, 38(2): 103.
[98]

Nacharaju P, Ko L W, Yen S H C. J. Neurochem., 1997, 69(4): 1709.
[99]

Necula M, Kuret J. J. Biol. Chem., 2004, 279(48): 49694.
[100]

Kuhla B, Haase C, Flach K, Lüth H J, Arendt T, Münch G. J. Biol. Chem., 2007, 282(10): 6984.
[101]

Yan S D, Chen X, Schmidt A M, Brett J, Godman G, Zou Y S, Scott C W, Caputo C, Frappier T, Smith M A. PNAS, 1994, 91(16): 7787.
[102]

Ko L W, Ko E C, Nacharaju P, Liu W K, Chang E, Kenessey A, Yen S H C. Brain Res., 1999, 830(2): 301.
[103]

Smith M A, Tabaton M, Perry G. Neurosci. Lett., 1996, 217(2/3): 210.
[104]

Ledesma M D, Medina M, Avila J. Mol. Chem. Neuropathol., 1996, 27(3): 249.
[105]

Zheng Y J, Thompson P R, Cebrat M, Wang L, Devlin M K, Alani R M, Cole P A.. Chromatin and Chromatin Remodeling Enzymes, Part B. Amsterdam: Elsevier, 2003, 376: 188.
[106]

Min S W, Cho S H, Zhou Y G, Schroeder S, Haroutunian V, Seeley W W, Huang E J, Shen Y, Masliah E, Mukherjee C, Meyers D, Cole P A, Ott M, Gan L. Neuron, 2010, 67(6): 953.
[107]

Cripps D, Thomas S N, Jeng Y, Yang F, Davies P, Yang A J. J. Biol. Chem., 2006, 281(16): 10825.
[108]

Paine S, Bedford L, Thorpe J R, Mayer R J, Cavey J R, Bajaj N, Sheppard P W, Lowe J, Layfield R. Neurosci. Lett., 2009, 460(3): 205.
[109]

Morishima-Kawashima M, Hasegawa M, Takio K, Suzuki M, Titani K, Ihara Y. Neuron, 1993, 10(6): 1151.
[110]

Petrucelli L, Dickson D, Kehoe K, Taylor J, Snyder H, Grover A, De Lucia M, McGowan E, Lewis J, Prihar G, Kim J, Dillmann W H, Browne S E, Hall A, Voellmy R, Tsuboi Y, Dawson T M, Wolozin B, Hardy J, Hutton M. Human Molecular Genetics, 2004, 13 (7): 703.
[111]

Shimura H, Schwartz D, Gygi S P, Kosik K S. J. Biol. Chem., 2004, 279(6): 4869.
[112]

Babu J R, Geetha T, Wooten M W. J. Neurochem., 2005, 94(1): 192.
[113]

Flach K, Ramminger E, Hilbrich I, Arsalan-Werner A, Albrecht F, Herrmann L, Goedert M, Arendt T, Holzer M. Biochim. Biophys. Acta BBA Mol. Basis Dis., 2014, 1842(9): 1527.
[114]

Keck S, Nitsch R, Grune T, Ullrich O. J. Neurochem., 2003, 85(1): 115.
[115]

LÓpez Salon M, Morelli L, Castaño E M, Soto E F, Pasquini J M. J. Neurosci. Res., 2000, 62(2): 302.
[116]

Ohshima H, Friesen M, Brouet I, Bartsch H. Food Chem. Toxicol., 1990, 28(9): 647.
[117]

Radi R, Peluffo G, Alvarez M N, Naviliat M, Cayota A. Free. Radic. Biol. Med., 2001, 30(5): 463.
[118]

Tsikas D, Duncan M W. Mass Spectrom. Rev., 2014, 33(4): 237.
[119]

Reynolds M R, Berry R W, Binder L I. Biochemistry, 2005, 44(42): 13997.
[120]

Reynolds M R, Berry R W, Binder L I. Biochemistry, 2005, 44(5): 1690.
[121]

Reyes J F, Reynolds M R, Horowitz P M, Fu Y F, Guillozet-Bongaarts A L, Berry R, Binder L I. Neurobiol. Dis., 2008, 31(2): 198.
[122]

Dorval V, Fraser P E. J. Biol. Chem., 2006, 281(15): 9919.
[123]

Takahashi K, Ishida M, Komano H, Takahashi H. Neurosci. Lett., 2008, 441(1): 90.
[124]

Schweers O, Mandelkow E M, Biernat J, Mandelkow E. PNAS, 1995, 92(18): 8463.
[125]

Zhou X Z, Kops O, Werner A, Lu P J, Shen M H, Stoller G, Küllertz G, Stark M, Fischer G, Lu K P. Mol. Cell, 2000, 6(4): 873.
[126]

Ugolini G, Cattaneo A, Novak M. NeuroReport, 1997, 8(17): 3709.
[127]

Prasanna Murthy S N, Wilson J H, Lukas T J, Kuret J, Lorand L. J. Neurochem., 1998, 71(6): 2607.
[128]

Luo H B, Xia Y Y, Shu X J, Liu Z C, Feng Y, Liu X H, Yu G, Yin G, Xiong Y S, Zeng K, Jiang J, Ye K Q, Wang X C, Wang J Z. PNAS, 2014, 111(46): 16586.
[129]

Landino L M, Skreslet T E, Alston J A. J. Biol. Chem., 2004, 279(33): 35101.
[130]

Bulbarelli A, Lonati E, Cazzaniga E, Gregori M, Masserini M. Mol. Cell. Neurosci., 2009, 42(1): 75.
[131]

Greenberg C S, Birckbichler P J, Rice R H. FASEB J., 1991, 5(15): 3071.
[132]

Singer S M, Zainelli G M, Norlund M A, Lee J M, Muma N A. Neurochem. Int., 2002, 40(1): 17.
[133]

Halverson R A, Lewis J, Frausto S, Hutton M, Muma N A. Journal of Neuroscience, 2005, 25 (5): 1226.
[134]

Du J T, Li Y M, Wei, Wu G S, Zhao Y F, Kanazawa K, Nemoto T, Nakanishi H. J. Am. Chem. Soc., 2005, 127(47): 16350.
[135]

Zhou L X, Zeng Z Y, Du J T, Zhao Y F, Li Y M. Biochem. Biophys. Res. Commun., 2006, 348(2): 637.
[136]

Du J T, Yu C H, Zhou L X, Wu W H, Lei P, Li Y, Zhao Y F, Nakanishi H, Li Y M. FEBS J., 2007, 274(19): 5012.
[137]

Brister M A, Pandey A K, Bielska A A, Zondlo N J. J. Am. Chem. Soc., 2014, 136(10): 3803.
[138]

Yu C H, Si T, Wu W H, Hu J, Du J T, Zhao Y F, Li Y M. Biochem. Biophys. Res. Commun., 2008, 375(1): 59.
[139]

Ma M R, Hu Z W, Zhao Y F, Chen Y X, Li Y M. Sci. Rep., 2016, 6: 37130.
[140]

Li Q Q, Liu Y Q, Luo Y Y, Chu T T, Gao N, Chen P G, Chen Y X, Li Y M. Chem. Commun., 2020, 56(40): 5370.
[141]

Zhao K, Lim Y J, Liu Z Y, Long H F, Sun Y P, Hu J J, Zhao C Y, Tao Y Q, Zhang X, Li D, Li Y M, Liu C. PNAS, 2020, 117(33): 20305.
[142]

Hu Z W, Ma M R, Chen Y X, Zhao Y F, Qiang W, Li Y M. J. Biol. Chem., 2017, 292(21): 2611.
[143]

Munari F, Barracchia C G, Franchin C, Parolini F, Capaldi S, Romeo A, Bubacco L, Assfalg M, Arrigoni G, D'Onofrio M. Angew. Chem. Int. Ed., 2020, 59(16): 6607.
[144]

Munari F, Barracchia C G, Parolini F, Tira R, Bubacco L, Assfalg M, D’Onofrio M. Int. J. Mol. Sci., 2020, 21(12): 4400.
[145]

Broncel M, Krause E, Schwarzer D, Hackenberger C P R. Chem. Eur. J., 2012, 18(9): 2488.
[146]

Ellmer D, Brehs M, Haj-Yahya M, Lashuel H A, Becker C F W. Angew. Chem. Int. Ed., 2019, 58(6): 1616.
[147]

Schwagerus S, Reimann O, Despres C, Smet-Nocca C, Hackenberger C P R. J. Pept. Sci., 2016, 22(5): 327.
[148]

Reynolds M R, Lukas T J, Berry R W, Binder L I. Biochemistry, 2006, 45(13): 4314.
[149]

Carlomagno Y, Chung D E C, Yue M, Castanedes-Casey M, Madden B J, Dunmore J, Tong J M, DeTure M, Dickson D W, Petrucelli L, Cook C. J. Biol. Chem., 2017, 292(37): 15277.
[150]

Rani L, Mittal J, Mallajosyula S S. J. Phys. Chem. B, 2020, 124(10): 1909.
[151]

Zhao D S, Chen Y X, Li Y M. Chem. Commun., 2015, 51(11): 2095.

Funding

National Key Research and Development Program of China(2018YFA0507600)

National Key Research and Development Program of China(2019YFA0904200)

National Natural Science Foundation of China(92053108)

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