蛋白质氨基酸残基及游离氨基酸的化学修饰与应用

doi:10.11949/0438-1157.20240707

摘要/Abstract

摘要：

蛋白质是细胞内含量丰富的生物分子之一，其参与几乎所有的生化过程。总结了蛋白质中氨基酸残基和游离氨基酸的反应类型，包括半胱氨酸的巯基通过烷基化、不饱和键加成、氟取代等反应类型与探针或试剂反应；赖氨酸的氨基与醛或酯发生缩合反应，还与烯键或炔键发生加成反应，少量可与双官能团底物发生环化反应；精氨酸的胍基更多的是通过与探针形成非共价氢键实现修饰；含羧基侧链的谷氨酸和天冬氨酸与不同类型的底物进行酯化和自由基等反应；酪氨酸的酚羟基和邻位C上的氢可发生自由基及亲核取代反应；含氮杂环的吲哚基（色氨酸）及咪唑基（组氨酸）有不同反应位点，可发生特定类型的反应。同时，对于蛋白质残基修饰在医药与工程领域的相关应用进行了概括。通过以上总结对不同氨基酸的修饰及应用提出不足和对未来的展望。

关键词: 蛋白质, 氨基酸残基, 游离氨基酸, 肽, 化学反应, 生物化学

Abstract:

Proteins are one of the abundant biomolecules in cells, which participate in almost all biochemical processes. The reaction types of amino acid residues and free amino acids (in vivo and/or in vitro) were summarized, such as alkylation, unsaturated bond addition and fluorinated substitution of cysteine sulfhydryl groups with probes or reagents. The amino group of lysine can undergo condensation reaction with aldehydes or esters, and can also reacts with alkene bonds or acetylene bonds as well as a few cases of cyclization with bi-functional substrates. To obtain its modification, the guanidine group of arginine prefers to form non-covalent hydrogen bonds with the probe. Different substrates, such as esterification and free radicals can modify the carboxyl side chains containing amino acids (glutamate and aspartate). The hydroxyl group of tyrosine phenol and the hydrogen at the ortho-site C can undergo free radical and nucleophilic substitution reactions. The indolyl (tryptophan) and imidazolyl (histidine) are typical nitrogen heterocyclic rings that can have specific types of reactions. Based on the above summary, we concluded the shortcomings and prospects of modification of different amino acids.

Key words: protein, amino acid residue, free amino acid, peptide, chemical reaction, biochemistry

中图分类号:

TQ 932

苗青, 石睿思. 蛋白质氨基酸残基及游离氨基酸的化学修饰与应用[J]. 化工学报, DOI: 10.11949/0438-1157.20240707.

Qing MIAO, Ruisi SHI. Chemical modification and applications of protein amino acid residues and free amino acids[J]. CIESC Journal, DOI: 10.11949/0438-1157.20240707.

图/表 14

图1 与巯基发生卤代烷基化反应的机理及相应探针或试剂

Fig.1 Mechanisms of haloalkylation reactions with sulfhydryl groups and corresponding probes or reagents（X=F,Cl,Br，螺旋结构为蛋白质）

图2 不饱和键与巯基的反应类型：（a）烯键与半胱氨酸巯基的加成反应；（b）锍中心引发的硫醇-炔型反应；（c）巯基与炔键的反应；（d）巯基与类马来酰亚胺的反应

Fig.2 Reaction types of unsaturated bonds with sulfhydryl groups:(a) the addition of alkene bond to cysteine sulfhydryl group; (b) thiol-alkyne type reactions initiated by sulfonium centres; (c) reaction of sulfhydryl groups with alkyne bonds; (d) reaction of sulfhydryl groups with maleimide analogues

图3 基于氟取代反应的机理及探针或试剂（红色标记为反应性氟，彩色带状结构代表蛋白质，绿色方块代表生物素，荧光团等功能基团）

Fig.3 Mechanism and probes or reagents based on fluorine substitution reactions

图4 与半胱氨酸残基的各种类型反应：（a）类似硫交换反应；（b）和（c）三元环类型；（d）、（e）和（f）其它类型反应

Fig.4 Various types of reactions with cysteine residues: (a) sulfur-like exchange reaction; (b) and (c) ternary ring types; (d), (e) and (f) other types of reactions

图5 游离半胱氨酸的修饰反应：（a）和（b）卤代烷基化反应；（c）和（d）硫交换反应；（e）和（f）形成五元环反应；（g）取代反应

Fig.5 Modification reactions of free cysteine: (a) and (b) haloalkylation reactions; (c) and (d) sulphur exchange reactions; (e) and (f) form a five-membered ring reaction; (g) substitution reactions

图6 丙烯酸酯的特殊不饱和键与巯基反应示意图和相应的探针结构

Fig.6 Schematic of the reaction of special unsaturated bond with sulfhydryl group of acrylate and corresponding probe structure（圆圈代表不同探针的丙烯酸酯外的其他部分）

图7 与赖氨酸残基的环化反应及与醛或酯的反应：（a）、（b）、（c）和（d）环化反应；（e）、（f）、（g）、（h）、（i）和（j）醛或酯的反应

Fig.7 Cyclisation reactions with lysine residues and reactions with aldehydes or esters: (a), (b), (c) and (d) cyclisation reactions; (e), (f), (g), (h), (i) and (j) aldehyde or ester reactions

图8 赖氨酸与烯（a）、炔（b）键的反应以及既可修饰游离赖氨酸又可修饰游离精氨酸的探针

Fig.8 Reactions of lysine with alkene (a) and alkynyl (b) bonds and probes that modify both free lysine and free arginine

图9 探针或试剂与游离赖氨酸的修饰：（a）和（b）质子化反应；（c）亲核取代反应；（d）环化反应；（e）加成反应

Fig.9 Modification of probes or reagents with free lysine: (a) and (b) protonation reactions; (c) nucleophilic substitution reactions; (d) cyclisation reactions; (e) addition reactions

图10 精氨酸的修饰：1-7为游离精氨酸的标记探针；8为蛋白质中精氨酸残基的标记探针

Fig.10 Arginine modification: 1-7 are labelled probes for free arginine; 8 is a labelling probe for arginine residues in proteins

图11 谷氨酸或天冬氨酸的羧基修饰分为四类：（a）通过Ynamide试剂反应；（b）、（c）、（d）与三元环反应；（e）、（f）与四唑的反应；（g）、（h）与重氮基团反应

Fig.11 Carboxyl modifications of glutamic acid or aspartic acid are divided into four categories: (a) reaction by Ynamide reagent; (b), (c), (d) reactions with the ternary ring; (e), (f) reactions with tetrazoles; (g), (h) reactions with diazo groups

图12 酪氨酸残基两种类型的修饰：1-6为修饰酪氨酸侧链酚羟基的探针；7-12为修饰与酪氨酸侧链酚羟基邻位C上H的探针

Fig.12 Two types of modifications of tyrosine residues: 1-6 are probes for modification of the phenolic hydroxyl group of the tyrosine side chain; 7-12 is the probe that modifies H on the ortho C of the phenol hydroxyl group of the side chain of tyrosine

图13 色氨酸通过吲哚环上的C-1、N-H、C-2、C-7位点及不同位点环的修饰

Fig.13 Tryptophan is modified by the C-1, N-H, C-2, and C-7 sites on the indole ring and different site-specific rings

图14 组氨酸不同位点的修饰（分别为通过N-1、C-2位点进行反应，以及氧化环化反应）

Fig.14 Modification of different sites of histidine (reactions through the N-1 and C-2 sites, and oxidative cyclisation, respectively)

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