N-磷酸化修饰蛋白质的富集和鉴定方法

doi:10.3724/SP.J.1123.2019.09015

摘要/Abstract

摘要：

蛋白质磷酸化修饰在细胞的信号转导、代谢、发育等生命过程中发挥着重要作用。除了研究较为透彻的发生在丝氨酸、苏氨酸和酪氨酸侧链羟基的O-磷酸化修饰之外，近年来，发生在组氨酸、精氨酸和赖氨酸侧链氨基的N-磷酸化修饰受到了越来越广泛的关注。然而，由于N-磷酸化修饰具有独特的P-N键结构，导致其化学稳定性差。尤其是在O-磷酸化肽段富集常用的酸性条件下，N-磷酸化极易丢失。因此，目前对N-磷酸化蛋白质的研究仍处于初始阶段。该文针对蛋白质N-磷酸化修饰的特点、富集和鉴定方法进行了综述，并对其发展前景进行了展望。

关键词: 蛋白质N-磷酸化修饰, 富集, 检测, 综述

Abstract:

Protein phosphorylation plays an important role in various life processes, such as signaling, metabolism, and growth. In addition to the well-known O-phosphorylation, which occurs on serine, threonine and tyrosine side chains, N-phosphorylation, occurring on histidine, arginine and lysine side chains, has attracted much attention. However, due to the chemical instability of N-phosphorylation, the studies on it are still in the nascent stage. In this review, enrichment and detection methods for protein N-phosphorylation are reviewed, and future research in this field is prospected.

Key words: protein N-phosphorylation, enrichment, detection, review

胡晔晨, 江波, 张丽华, 张玉奎. N-磷酸化修饰蛋白质的富集和鉴定方法[J]. 色谱, 2020, 38(3): 278-286.

HU Yechen, JIANG Bo, ZHANG Lihua, ZHANG Yukui. Advancesin enrichment and detection methods for N-phosphorylated proteins[J]. Chinese Journal of Chromatography, 2020, 38(3): 278-286.

图/表 6

图1 蛋白质N-磷酸化修饰的化学结构

Fig. 1 Chemical structures of protein N-phosphorylation

图2 (a) pHis的类似物与(b)3种合成肽文库的结构, 其中His或稳定的pHis类似物(6或7)被随机地插入中性氨基酸(丙氨酸和甘氨酸)中

Fig. 2 (a)Analogues of pHis and (b) structures of the three synthetic peptide libraries used in which either His or a stable pHis mimetic (6 or 7) is flanked by randomized, neutral amino acids (alanine and glycine)

图3 pHis肽段的富集和鉴定流程图[41]

Fig. 3 Flow chart of the enrichment and identification of pHis peptides[41] CID: collision induced dissociation.

图4 抗pArg抗体的开发策略[45]

Fig. 4 Strategy to develop anti-pArg antibodies[45] KLH: keyhole limpet hemocyanin.

图5 (a) ReDD策略的流程图和(b)pLys肽段发生的化学过程[52]

Fig. 5 (a) Flowchart of retention time difference combining dimethyl labeling (ReDD) strategy and (b) chemical process of pLys peptide[52] FA: formic acid; SCX: strong cation exchange chromatography.

图6 (a) CHD策略的流程图和(b)pLys肽段发生的化学过程[53]

Fig. 6 (a) Flowchart of cleavable hydrophobic derivatization (CHD) strategy and (b) chemical process of pLys peptide[53] Dec-disulf-NHS: 2, 5-dioxopyrrolidin-1-yl-3-(decyldisulfanyl)propanoate; TCEP: tris(2-carboxyethyl)phosphine.

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