3D Structure of the Transient Intermediate of the Enzyme-Substrate Complex of Sortase A Reveals How Calcium Binding and Substrate Recognition Cooperate in Substrate Activation

被引：7

作者：

Chen, Jia-Liang ^{[1
,2
]}

Wang, Xiao ^{[1
]}

Yang, Feng ^{[1
]}

Li, Bin ^{[1
]}

Otting, Gottfried ^{[3
]}

Su, Xun-Cheng ^{[1
]}

机构：

[1] Nankai Univ, Coll Chem, State Key Lab Elemento Organ Chem, Tianjin Key Lab Biosensing & Mol Recognit, Tianjin 300071, Peoples R China

[2] Zaozhuang Univ, Coll Chem Chem Engn & Mat Sci, Zaozhuang 277160, Shandong, Peoples R China

[3] Australian Natl Univ, Australian Res Council Ctr Excellence Innovat Pept, Res Sch Chem, Canberra 2601, Australia

来源：

ACS CATALYSIS | 2023年 / 13卷 / 17期

基金：

中国国家自然科学基金; 澳大利亚研究理事会;

关键词：

enzyme; structure of enzyme intermediate; unstableprotein complex; paramagnetic NMR; protein labeling; GENERAL FORCE-FIELD; STAPHYLOCOCCUS-AUREUS; A TRANSPEPTIDASE; SURFACE-PROTEINS; SORTING SIGNAL; XPLOR-NIH; CELL-WALL; PROTEASE; SERINE; SPECIFICITY;

D O I：

10.1021/acscatal.3c02214

中图分类号：

O64 [物理化学（理论化学）、化学物理学];

学科分类号：

070304 ; 081704 ;

摘要：

We report the solution structure of an authentic sortaseA thioesterintermediate, including the bound substrate. The structure was determinedby a combination of site-specific tagging, selective isotope labeling,and paramagnetic nuclear magnetic resonance spectroscopy. Pseudocontactshifts generated with different lanthanide tags delivered the conformationof the isotope-labeled bound substrate at atomic resolution, enablingthe determination of the complete 3D structure of the short-livedand lowly populated enzymatic reaction intermediate in solution. Theformation of the unstable thioester complex is accompanied by an increasein calcium binding affinity. Furthermore, the intermediate is significantlystabilized by calcium and decays quickly upon removal of calcium.Cooperativity between calcium binding and substrate recognition thusplays an important role in the function of sortase A.

引用

页码：11610 / 11624

页数：15

共 72 条

[1] Energetic dissection of Gleevec's selectivity toward human tyrosine kinases [J].