Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/96231
Citations
Scopus Web of Science® Altmetric
?
?
Type: Journal article
Title: Mechanisms of inhibition of Rhizobium etli pyruvate carboxylase by L-Aspartate
Author: Sirithanakorn, C.
Adina-Zada, A.
Wallace, J.
Jitrapakdee, S.
Attwood, P.
Citation: Biochemistry, 2014; 53(45):7100-7106
Publisher: ACS
Issue Date: 2014
ISSN: 0006-2960
1520-4995
Statement of
Responsibility: 
Chaiyos Sirithanakorn, Abdussalam Adina-Zada, John C. Wallace, Sarawut Jitrapakdee, and Paul V. Attwood
Abstract: L-aspartate is a regulatory feedback inhibitor of the biotin-dependent enzyme pyruvate carboxylase in response to increased levels of tricarboxylic acid cycle intermediates. Detailed studies of L-aspartate inhibition of pyruvate carboxylase have been mainly confined to eukaryotic microbial enzymes, and aspects of its mode of action remain unclear. Here we examine its inhibition of the bacterial enzyme Rhizobium etli pyruvate carboxylase. Kinetic studies demonstrated that L-aspartate binds to the enzyme cooperatively and inhibits the enzyme competitively with respect to acetyl-CoA. L-aspartate also inhibits activation of the enzyme by MgTNP-ATP. The action of L-aspartate was not confined to inhibition of acetyl-CoA binding, because the acetyl-CoA-independent activity of the enzyme was also inhibited by increasing concentrations of L-aspartate. This inhibition of acetyl-CoA-independent activity was demonstrated to be focused in the biotin carboxylation domain of the enzyme, and it had no effect on the oxamate-induced oxaloacetate decarboxylation reaction that occurs in the carboxyl transferase domain. L-aspartate was shown to competitively inhibit bicarbonate-dependent MgATP cleavage with respect to MgATP but also probably inhibits carboxybiotin formation and/or translocation of the carboxybiotin to the site of pyruvate carboxylation. Unlike acetyl-CoA, L-aspartate has no effect on the coupling between MgATP cleavage and oxaloacetate formation. The results suggest that the three allosteric effector sites (acetyl-CoA, MgTNP-ATP, and L-aspartate) are spatially distinct but connected by a network of allosteric interactions.
Keywords: Rhizobium etli
Pyruvate Carboxylase
Aspartic Acid
Enzyme Inhibitors
Rights: This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes. © 2014 American Chemical Society
DOI: 10.1021/bi501113u
Appears in Collections:Aurora harvest 3
Molecular and Biomedical Science publications

Files in This Item:
File Description SizeFormat 
hdl_96231.pdfPublished version520.61 kBAdobe PDFView/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.