基于化学蛋白质组学的激酶组学研究进展

蛋白激酶是细胞内信号转导通路网络的关键组成部分，调节细胞生长、分化、代谢及生存等重要生物学过程。激酶组（kinome）是指细胞或组织中所有激酶的总称。激酶组学（kinomics）是研究激酶组的学科，包括激酶的丰度、活性、底物特异性、磷酸化状态及氨基酸突变。目前人激酶组由568种基因编码组成，其中约50%激酶定位于疾病基因位点。由于基因扩增或突变引起的蛋白激酶活性失调与许多疾病的发生相关，包括炎症、糖尿病及各种肿瘤，因此，人激酶组被认为是药物靶点的潜在资源库。靶向激酶的小分子抑制剂已成功用于肿瘤治疗。化学蛋白质组技术是一种新的深入研究激酶组的方法，是将亲和富集与质谱技术相结合，在生理状态下研究激酶丰度及磷酸化水平。通常情况下，将一个或多个广谱小分子激酶抑制剂偶联于固相载体（如琼脂糖凝胶），亲和富集细胞或组织中的所有激酶，然后通过质谱鉴定或定量。该方法可用于激酶抑制剂药物或药物候选物的靶点专一性及耐药机制研究，为深入研究药物作用机制及寻找联合用药新靶点提供依据。对人激酶组及肿瘤激酶组无偏见、全景式分析能帮助发现更多新药靶点，以及激酶活性谱与肿瘤治疗的相关性，为个体化治疗提供依据。本文对人激酶组、激酶、激酶抑制剂与肿瘤的关系，基于化学蛋白质组的激酶组学研究进展及其在药物研究中的最新应用进行综述。

Progress in chemical proteomics-based kinome study

Protein kinases are key components of cell signaling networks and thereby regulate fundamental biological processes such as cellular growth, proliferation, metabolism and survival. Kinome refers to all kinases in cells or tissue and “kinomics”is the global analysis of kinome with respect to abundance, activity, substrate specificity, phosphorylation pattern and mutational status. Human kinome currently contains 568 members, nearly half of which can be mapped to disease loci and deregulation of kinase activity by gene amplifica-tion or mutations has been implicated in diseases such as inflammation, diabetes and cancer. Therefore, human kinome is being recognized as a potentially rich source of drug targets. Kinase inhibitors have been successfully used to treat many kinds of advanced cancers. Chemi-cal proteomics is emerging as a novel comprehensive kinome approach that combines an immobilized inhibitor affinity pull-down approach with mass spectrometry-based proteomics for kinase identification, quantification and phosphorylation analysis under physiological condi-tion. Commonly, one or multiple broad-spectrum kinase inhibitors are covalently immobilized on a biocompatible matrix such as sepharose to enrich all kinases in cells or tissue and then the kinases are identified and quantified by mass spectrometry analysis. It can be used to study the specificity of kinase inhibitor drug, drug candidate or drug resistance mechanism, which can help to understand the mechanism and find combinational drug target. Large-scale unbiased kinome and cancer kinome study will facilitate new drug target discovery and correlate tumor tissue kinome profiles with response to therapy and therefore may be used for future therapy selection in personalized medicine. In this paper, the human kinome, kinase, kinase inhibitor and cancer, chemical proteomics based kinome study progress and its applications in drug discovery are reviewed.