Determination of the binding specificity of the 12S subunit of the transcarboxylase by saturation transfer difference NMR.
In this study we present the characterization of the interaction of biotin and methylmalonyl-CoA (MMCoA) with the carboxyltransferase subunit (12S) from the transcarboxylase (TC) from Propionibacterium shermanii. This biotin dependent multienzyme complex catalyses the transfer of carbon dioxide from methylmalonyl-CoA (MMCoA) to pyruvate. The Saturation Transfer Difference NMR (STD) technique was performed to determine the binding epitope from biotin and MMCoA to the 12S subunit. We could show by titrations during STD experiments that biotin and MMCoA bind cooperatively in one binding pocket.
Peikert
Claudia
C
Institute of Analytical Chemistry, University of Leipzig, Linnestr. 3, D-04103 Leipzig, Germany.
Seeger
Karsten
K
Bhat
Rakesh Kumar
RK
Berger
Stefan
S
D000214:Acyl Coenzyme A / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D001710:Biotin / Q000378:metabolism; D019878:Carboxyl and Carbamoyl Transferases / Q000737:chemistry / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011424:Propionibacterium / Q000201:enzymology; D011485:Protein Binding; D021122:Protein Subunits; D013379:Substrate Specificity; D017186:Titrimetry / Q000379:methods
D000214:Acyl Coenzyme A; D021122:Protein Subunits; C015357:methylmalonyl-coenzyme A; D001710:Biotin; D019878:Carboxyl and Carbamoyl Transferases; C021580:Methylmalonyl-CoA carboxytransferase