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PMID: 15329673 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Transcarboxylase 5S structures: assembly and catalytic mechanism of a multienzyme complex subunit.

The EMBO journal ·Vol. 23 ·No. 18 ·2004-09-15 ·Pages 3621-31

Hall PR, Zheng R, Antony L, Pusztai-Carey M, Carey PR, Yee VC

Abstract

Transcarboxylase is a 1.2 million Dalton (Da) multienzyme complex from Propionibacterium shermanii that couples two carboxylation reactions, transferring CO(2)(-) from methylmalonyl-CoA to pyruvate to yield propionyl-CoA and oxaloacetate. Crystal structures of the 5S metalloenzyme subunit, which catalyzes the second carboxylation reaction, have been solved in free form and bound to its substrate pyruvate, product oxaloacetate, or inhibitor 2-ketobutyrate. The structure reveals a dimer of beta(8)alpha(8) barrels with an active site cobalt ion coordinated by a carbamylated lysine, except in the oxaloacetate complex in which the product's carboxylate group serves as a ligand instead. 5S and human pyruvate carboxylase (PC), an enzyme crucial to gluconeogenesis, catalyze similar reactions. A 5S-based homology model of the PC carboxyltransferase domain indicates a conserved mechanism and explains the molecular basis of mutations in lactic acidemia. PC disease mutations reproduced in 5S result in a similar decrease in carboxyltransferase activity and crystal structures with altered active sites.

MeSH Terms
Amino Acid Sequence Bacterial Proteins/chemistry,genetics,isolation & purification Binding Sites Butyrates/metabolism Carboxyl and Carbamoyl Transferases/chemistry,genetics,isolation & purification Catalytic Domain Crystallography, X-Ray Enzyme Inhibitors/metabolism Humans Molecular Sequence Data Multienzyme Complexes/chemistry Mutagenesis, Site-Directed Oxaloacetic Acid/metabolism Propionibacterium/enzymology Protein Binding Protein Conformation Protein Subunits Pyruvate Carboxylase/chemistry,genetics,metabolism Pyruvic Acid/metabolism Sequence Homology, Amino Acid
Chemicals
Bacterial Proteins Butyrates Enzyme Inhibitors Multienzyme Complexes Protein Subunits Oxaloacetic Acid alpha-ketobutyric acid Pyruvic Acid Carboxyl and Carbamoyl Transferases Methylmalonyl-CoA carboxytransferase Pyruvate Carboxylase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Hall Pamela R
Department of Pharmacology, Case Western Reserve University, Cleveland, OH, USA.
Zheng Run
Antony Lizamma
Pusztai-Carey Marianne
Carey Paul R
Yee Vivien C
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7600373
Published
2004-09-15
Epub
2004-00-26
Pages
3621-31
Language
English
Region
England
NLM ID
8208664
PMCID
PMC517613
Subset
IM
Grants
NIDDK NIH HHS · R01 DK053053 · United States
NIGMS NIH HHS · T32 GM008803 · United States
NIDDK NIH HHS · DK053053 · United States
NIGMS NIH HHS · GM08803 · United States
Databases
PDB
1RQB, 1RQE, 1RQH, 1RR2, 1S3H, 1U5J
External Links
PubMed source DOI: 10.1038/sj.emboj.7600373 PMC Full Text View journal details
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