Centre for Cancer Research, Monash Institute of Medical Research, Melbourne, Victoria, Australia.
Oncogene. 2011 Oct 27;30(43):4375-85. doi: 10.1038/onc.2011.177. Epub 2011 May 23.
Integrin-linked kinase (ILK) is a highly evolutionarily conserved intracellular protein that was originally identified as an integrin-interacting protein, and extensive genetic and biochemical studies have shown that ILK expression is vital during both embryonic development and tissue homeostasis. At the cellular and tissue levels, ILK regulates signaling pathways for cell adhesion-mediated cell survival (anoikis), apoptosis, proliferation and mitosis, migration, invasion, and vascularization and tumor angiogenesis. ILK also has central roles in cardiac and smooth-muscle contractility, and ILK dysregulation causes cardiomyopathies in humans. ILK protein levels are increased in several human cancers and often the expression level predicts poor patient outcome. Abundant evidence has accumulated suggesting that, of the diverse functions of ILK, some may require kinase activity whereas others depend on protein-protein interactions and are, therefore, independent of kinase activity. However, the past several years have seen an ongoing debate about whether ILK indeed functions as a protein serine/threonine kinase. This debate centers on the atypical protein kinase domain of ILK, which lacks some amino-acid residues thought to be essential for phosphotransferase activity. However, similar deficiencies are present in the catalytic domains of other kinases now known to possess protein kinase activity. Numerous studies have shown that ILK phosphorylates peptide substrates in vitro, corresponding to ILK-mediated phosphorylations in intact cells, and a recent report characterizing in vitro phosphotransferase activity of highly purified, full-length ILK, accompanied by detailed enzyme kinetic analyses, shows that, at least in vitro, ILK is a bona fide protein kinase. However, several genetic studies suggest that, not all biological functions of ILK require kinase activity, and that it can function as an adaptor/scaffold protein. Here, we review evidence for and against ILK being an active kinase, and provide a framework for strategies to further analyze the kinase and adaptor functions of ILK in different cellular contexts.
整合素连接激酶 (ILK) 是一种高度进化保守的细胞内蛋白,最初被鉴定为整合素相互作用蛋白,广泛的遗传和生化研究表明,ILK 的表达在胚胎发育和组织稳态中至关重要。在细胞和组织水平上,ILK 调节细胞黏附介导的细胞存活 (失巢凋亡)、细胞凋亡、增殖和有丝分裂、迁移、侵袭以及血管生成和肿瘤血管生成的信号通路。ILK 在心肌和平滑肌收缩中也具有核心作用,ILK 失调会导致人类心肌病。ILK 蛋白水平在几种人类癌症中增加,并且表达水平通常预测患者预后不良。大量证据表明,ILK 的多种功能中,有些可能需要激酶活性,而其他功能则依赖于蛋白-蛋白相互作用,因此不依赖于激酶活性。然而,过去几年一直在争论 ILK 是否确实作为一种丝氨酸/苏氨酸蛋白激酶发挥作用。这场争论的焦点是 ILK 的非典型蛋白激酶结构域,该结构域缺乏一些氨基酸残基,这些残基被认为对磷酸转移酶活性至关重要。然而,现在已知具有蛋白激酶活性的其他激酶的催化结构域也存在类似的缺陷。许多研究表明,ILK 在体外可磷酸化肽底物,与完整细胞中 ILK 介导的磷酸化相对应,最近一项关于高度纯化全长 ILK 体外磷酸转移酶活性的报告,并伴有详细的酶动力学分析,表明至少在体外,ILK 是一种真正的蛋白激酶。然而,一些遗传研究表明,并非 ILK 的所有生物学功能都需要激酶活性,并且它可以作为衔接子/支架蛋白发挥作用。在这里,我们回顾了支持和反对 ILK 作为活性激酶的证据,并提供了一个框架,用于进一步分析不同细胞环境中 ILK 的激酶和衔接子功能的策略。