Division of Medicinal Chemistry and Microbiology, Faculty of Chemistry, Wroclaw University of Technology, Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland.
Biochimie. 2012 Mar;94(3):704-10. doi: 10.1016/j.biochi.2011.10.014. Epub 2011 Nov 7.
Methionyl aminopeptidases (MetAPs) are metallo-dependent proteases responsible for removing of N-terminal methionine residue of peptides and proteins during protein maturation and activation. In this report we use a comprehensive strategy to screen the substrate specificity of three methionyl aminopeptidases: Homo sapiens MetAP-1, Homo sapiens MetAP-2 and Escherichia coli MetAP-1. By utilizing a 65-membered fluorogenic substrate library consisting of natural and unnatural amino acids we established detailed substrate preferences of each enzyme in the S1 pocket. Our results show that this pocket is highly conserved for all investigated MetAPs, very stringent for methionine, and that several unnatural amino acids with methionine-like characteristics were also well hydrolyzed by MetAPs. The substrate-derived results were verified using several phosphonate and phosphinate-based inhibitors.
甲硫氨酰氨基肽酶（MetAPs）是金属依赖性蛋白酶，负责在蛋白质成熟和激活过程中去除肽和蛋白质的 N 端甲硫氨酸残基。在本报告中，我们使用综合策略筛选了三种甲硫氨酰氨基肽酶的底物特异性：人源 MetAP-1、人源 MetAP-2 和大肠杆菌 MetAP-1。通过利用由天然和非天然氨基酸组成的 65 成员荧光底物文库，我们在 S1 口袋中建立了每种酶的详细底物偏好。我们的结果表明，该口袋在所有研究的 MetAPs 中高度保守，对甲硫氨酸非常严格，并且一些具有甲硫氨酸样特征的非天然氨基酸也被 MetAPs 很好地水解。使用几种膦酸酯和膦酸盐基抑制剂验证了基于底物的结果。
