Structure, mechanism and regulation of pyruvate carboxylase.
PC (pyruvate carboxylase) is a biotin-containing enzyme that catalyses the HCO(3)(-)- and MgATP-dependent carboxylation of pyruvate to form oxaloacetate. This is a very important anaplerotic reaction, replenishing oxaloacetate withdrawn from the tricarboxylic acid cycle for various pivotal biochemical pathways. PC is therefore considered as an enzyme that is crucial for intermediary metabolism, controlling fuel partitioning toward gluconeogenesis or lipogenesis and in insulin secretion. The enzyme was discovered in 1959 and over the last decade there has been much progress in understanding its structure and function. PC from most organisms is a tetrameric protein that is allosterically regulated by acetyl-CoA and aspartate. High-resolution crystal structures of the holoenzyme with various ligands bound have recently been determined, and have revealed details of the binding sites and the relative positions of the biotin carboxylase, carboxyltransferase and biotin carboxyl carrier domains, and also a unique allosteric effector domain. In the presence of the allosteric effector, acetyl-CoA, the biotin moiety transfers the carboxy group between the biotin carboxylase domain active site on one polypeptide chain and the carboxyltransferase active site on the adjacent antiparallel polypeptide chain. In addition, the bona fide role of PC in the non-gluconeogenic tissues has been studied using a combination of classical biochemistry and genetic approaches. The first cloning of the promoter of the PC gene in mammals and subsequent transcriptional studies reveal some key cognate transcription factors regulating tissue-specific expression. The present review summarizes these advances and also offers some prospects in terms of future directions for the study of this important enzyme.
Jitrapakdee
Sarawut
S
Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
St Maurice
Martin
M
Rayment
Ivan
I
Cleland
W Wallace
WW
Wallace
John C
JC
Attwood
Paul V
PV
D000105:Acetyl Coenzyme A / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D019731:Carbon-Nitrogen Ligases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D011485:Protein Binding; D011487:Protein Conformation; D011766:Pyruvate Carboxylase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D001710:Biotin; D000105:Acetyl Coenzyme A; D019731:Carbon-Nitrogen Ligases; C019791:biotin carboxylase; D011766:Pyruvate Carboxylase