The high-resolution structure of pig heart succinyl-CoA:3-oxoacid coenzyme A transferase.
The enzyme succinyl-CoA:3-oxoacid coenzyme A transferase (SCOT) participates in the metabolism of ketone bodies in extrahepatic tissues. It catalyses the transfer of coenzyme A (CoA) from succinyl-CoA to acetoacetate with a classical ping-pong mechanism. There is biochemical evidence that the enzyme undergoes conformational changes during the reaction, but no domain movements have been reported in the available crystal structures. Here, a structure of pig heart SCOT refined at 1.5 A resolution is presented, showing that one of the four enzyme subunits in the crystallographic asymmetric unit has a molecule of glycerol bound in the active site; the glycerol molecule is hydrogen bonded to the conserved catalytic glutamate residue and is likely to occupy the cosubstrate-binding site. The binding of glycerol is associated with a substantial relative movement (a 13 degrees rotation) of two previously undefined domains that close around the substrate-binding site. The binding orientation of one of the cosubstrates, acetoacetate, is suggested based on the glycerol binding and the possibility that this dynamic domain movement is of functional importance is discussed.
Coker
Shu-Fen
SF
Center for Amyloidosis and Acute Phase Proteins, Division of Medicine (Royal Free Campus), University College London, London, England.
Lloyd
Adrian J
AJ
Mitchell
Edward
E
Lewis
Gareth R
GR
Coker
Alun R
AR
Shoolingin-Jordan
Peter M
PM
D000595:Amino Acid Sequence; D000818:Animals; D015256:Coenzyme A-Transferases / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D005990:Glycerol / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009206:Myocardium / Q000201:enzymology*; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D013379:Substrate Specificity; D013552:Swine / Q000378:metabolism
D021122:Protein Subunits; D015256:Coenzyme A-Transferases; C021460:3-ketoacid CoA-transferase; D005990:Glycerol