2VJK

Formyl-CoA transferase with aspartyl-CoA thioester intermediate derived from oxalyl-CoA

Summary for 2VJK

• Entry DOI: 10.2210/pdb2vjk/pdb
• Related: 1P5H 1P5R 1T3Z 1T4C 1VGQ 1VGR 2VJL 2VJM 2VJN 2VJO 2VJP 2VJQ
• Descriptor: FORMYL-COENZYME A TRANSFERASE, COENZYME A, CHLORIDE ION, ... (5 entities in total)
• Functional Keywords: cytoplasm, transferase, class iii coa transferase
• Biological source: OXALOBACTER FORMIGENES
• Cellular location: Cytoplasm: O06644
• Total number of polymer chains: 2
• Total formula weight: 96391.45

Authors

Berthold, C.L.,Toyota, C.G.,Richards, N.G.J.,Lindqvist, Y. (deposition date: 2007-12-11, release date: 2007-12-25, Last modification date: 2024-11-20)

Primary citation

Berthold, C.L.,Toyota, C.G.,Richards, N.G.J.,Lindqvist, Y.
Reinvestigation of the Catalytic Mechanism of Formyl-Coa Transferase, a Class III Coa-Transferase.
J.Biol.Chem., 283:6519-, 2008

PubMed Abstract: Formyl-coenzyme A transferase from Oxalobacter formigenes belongs to the Class III coenzyme A transferase family and catalyzes the reversible transfer of a CoA carrier between formyl-CoA and oxalate, forming oxalyl-CoA and formate. Formyl-CoA transferase has a unique three-dimensional fold composed of two interlaced subunits locked together like rings of a chain. We here present an intermediate in the reaction, formyl-CoA transferase containing the covalent beta-aspartyl-CoA thioester, adopting different conformations in the two active sites of the dimer, which was identified through crystallographic freeze-trapping experiments with formyl-CoA and oxalyl-CoA in the absence of acceptor carboxylic acid. The formation of the enzyme-CoA thioester was also confirmed by mass spectrometric data. Further structural data include a trapped aspartyl-formyl anhydride protected by a glycine loop closing down over the active site. In a crystal structure of the beta-aspartyl-CoA thioester of an inactive mutant variant, oxalate was found bound to the open conformation of the glycine loop. Together with hydroxylamine trapping experiments and kinetic as well as mutagenesis data, the structures of these formyl-CoA transferase complexes provide new information on the Class III CoA-transferase family and prompt redefinition of the catalytic steps and the modified reaction mechanism of formyl-CoA transferase proposed here.

PubMed: 18162462
DOI: 10.1074/JBC.M709353200

Experimental method

X-RAY DIFFRACTION (1.97 Å)