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Title: A Symmetrical Tetramer for S. aureus Pyruvate Carboxylase in Complex with Coenzyme A

Abstract

Pyruvate carboxylase (PC) is a conserved metabolic enzyme with important cellular functions. We report crystallographic and cryo-electron microscopy (EM) studies of Staphylococcus aureus PC (SaPC) in complex with acetyl-CoA, an allosteric activator, and mutagenesis, biochemical, and structural studies of the biotin binding site of its carboxyltransferase (CT) domain. The disease-causing A610T mutation abolishes catalytic activity by blocking biotin binding to the CT active site, and Thr908 might play a catalytic role in the CT reaction. The crystal structure of SaPC in complex with CoA reveals a symmetrical tetramer, with one CoA molecule bound to each monomer, and cryo-EM studies confirm the symmetrical nature of the tetramer. These observations are in sharp contrast to the highly asymmetrical tetramer of Rhizobium etli PC in complex with ethyl-CoA. Our structural information suggests that acetyl-CoA promotes a conformation for the dimer of the biotin carboxylase domain of PC that might be catalytically more competent.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
Doe - Office Of Science
OSTI Identifier:
979990
Report Number(s):
BNL-92908-2010-JA
TRN: US201015%%1375
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Journal Name:
Structure
Additional Journal Information:
Journal Volume: 17; Journal Issue: 6
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE; 59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; BIOTIN; CARBOXYLASE; CHANNELING; COENZYMES; CRYSTAL STRUCTURE; DIMERS; ENZYMES; FUNCTIONS; INFORMATION; MICROSCOPY; MOLECULES; MUTAGENESIS; MUTATIONS; RHIZOBIUM; STAPHYLOCOCCUS; national synchrotron light source

Citation Formats

Yu, L, Xiang, S, Lasso, G, Gil, D, Valle, M, and Tong, L. A Symmetrical Tetramer for S. aureus Pyruvate Carboxylase in Complex with Coenzyme A. United States: N. p., 2009. Web. doi:10.1016/j.str.2009.04.008.
Yu, L, Xiang, S, Lasso, G, Gil, D, Valle, M, & Tong, L. A Symmetrical Tetramer for S. aureus Pyruvate Carboxylase in Complex with Coenzyme A. United States. https://doi.org/10.1016/j.str.2009.04.008
Yu, L, Xiang, S, Lasso, G, Gil, D, Valle, M, and Tong, L. Thu . "A Symmetrical Tetramer for S. aureus Pyruvate Carboxylase in Complex with Coenzyme A". United States. https://doi.org/10.1016/j.str.2009.04.008.
@article{osti_979990,
title = {A Symmetrical Tetramer for S. aureus Pyruvate Carboxylase in Complex with Coenzyme A},
author = {Yu, L and Xiang, S and Lasso, G and Gil, D and Valle, M and Tong, L},
abstractNote = {Pyruvate carboxylase (PC) is a conserved metabolic enzyme with important cellular functions. We report crystallographic and cryo-electron microscopy (EM) studies of Staphylococcus aureus PC (SaPC) in complex with acetyl-CoA, an allosteric activator, and mutagenesis, biochemical, and structural studies of the biotin binding site of its carboxyltransferase (CT) domain. The disease-causing A610T mutation abolishes catalytic activity by blocking biotin binding to the CT active site, and Thr908 might play a catalytic role in the CT reaction. The crystal structure of SaPC in complex with CoA reveals a symmetrical tetramer, with one CoA molecule bound to each monomer, and cryo-EM studies confirm the symmetrical nature of the tetramer. These observations are in sharp contrast to the highly asymmetrical tetramer of Rhizobium etli PC in complex with ethyl-CoA. Our structural information suggests that acetyl-CoA promotes a conformation for the dimer of the biotin carboxylase domain of PC that might be catalytically more competent.},
doi = {10.1016/j.str.2009.04.008},
url = {https://www.osti.gov/biblio/979990}, journal = {Structure},
number = 6,
volume = 17,
place = {United States},
year = {2009},
month = {1}
}