Residues in the acetyl CoA binding site of pyruvate carboxylase involved in allosteric regulation

Kamonman Choosangtong; Chaiyos Sirithanakorn; Abdul Adina-Zada; John C Wallace; Sarawut Jitrapakdee; Paul V Attwood

doi:10.1016/j.febslet.2015.06.034

Residues in the acetyl CoA binding site of pyruvate carboxylase involved in allosteric regulation

FEBS Lett. 2015 Jul 22;589(16):2073-9. doi: 10.1016/j.febslet.2015.06.034. Epub 2015 Jul 3.

Authors

Kamonman Choosangtong¹, Chaiyos Sirithanakorn², Abdul Adina-Zada³, John C Wallace⁴, Sarawut Jitrapakdee⁵, Paul V Attwood⁶

Affiliations

¹ Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand. Electronic address: kamonman.c@hotmail.com.
² Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand. Electronic address: yozinomycetes@hotmail.com.
³ School of Chemistry and Biochemistry, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia. Electronic address: adinazada@bigpond.com.
⁴ School of Molecular and Biomedical Sciences, University of Adelaide, Adelaide, SA 5005, Australia. Electronic address: john.wallace@adelaide.edu.au.
⁵ Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand. Electronic address: sarawut.jit@mahidol.ac.th.
⁶ School of Chemistry and Biochemistry, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia. Electronic address: paul.attwood@uwa.edu.au.

PMID: 26149215
DOI: 10.1016/j.febslet.2015.06.034

Abstract

We have examined the roles of Asp1018, Glu1027, Arg469 and Asp471 in the allosteric domain of Rhizobium etli pyruvate carboxylase. Arg469 and Asp471 interact directly with the allosteric activator acetyl coenzyme A (acetyl CoA) and the R469S and R469K mutants showed increased enzymic activity in the presence and absence of acetyl CoA, whilst the D471A mutant exhibited no acetyl CoA-activation. E1027A, E1027R and D1018A mutants had increased activity in the absence of acetyl CoA, but not in its presence. These results suggest that most of these residues impose restrictions on the structure and/or dynamics of the enzyme to affect activity.

Keywords: Acetyl coenzyme A; Allosteric regulation; Pyruvate carboxylase; Rhizobium etli.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Acetyl Coenzyme A / chemistry
Acetyl Coenzyme A / metabolism*
Adenosine Triphosphate / chemistry
Adenosine Triphosphate / metabolism
Allosteric Regulation
Allosteric Site
Amino Acid Substitution
Arginine / chemistry
Aspartic Acid / chemistry
Bacterial Proteins / chemistry
Bacterial Proteins / genetics
Bacterial Proteins / metabolism*
Bicarbonates / chemistry
Biocatalysis
Glutamic Acid / chemistry
Kinetics
Magnesium / chemistry
Models, Molecular*
Molecular Conformation
Mutagenesis, Site-Directed
Mutant Proteins / chemistry
Mutant Proteins / metabolism
Protein Stability
Pyruvate Carboxylase / chemistry
Pyruvate Carboxylase / genetics
Pyruvate Carboxylase / metabolism*
Pyruvic Acid / chemistry
Pyruvic Acid / metabolism
Recombinant Proteins / chemistry
Recombinant Proteins / metabolism
Rhizobium etli / enzymology*
Rhizobium etli / metabolism

Substances

Bacterial Proteins
Bicarbonates
Mutant Proteins
Recombinant Proteins
Aspartic Acid
Glutamic Acid
Acetyl Coenzyme A
Pyruvic Acid
Adenosine Triphosphate
Arginine
Pyruvate Carboxylase
Magnesium

Grants and funding

GM070455/GM/NIGMS NIH HHS/United States