Determination of the Solution Structure of Malonyl-CoA by Two-Dimensional Nuclear Magnetic Resonance Spectroscopy and Dynamical Simulated Annealing Calculations

  • Jung, Jin-Won (Department of Biochemistry, College of Science, Yonsei University) ;
  • An, Jae-Hyung (Department of Biochemistry, College of Science, Yonsei University) ;
  • Kim, Yu-Sam (Department of Biochemistry, College of Science, Yonsei University) ;
  • Bang, Eun-Jung (Korea Basic Science Institute) ;
  • Lee, Weon-Tae (Department of Biochemistry, College of Science, Yonsei University)
  • Received : 1999.01.29
  • Accepted : 1999.03.18
  • Published : 1999.05.31

Abstract

In order to understand the initial interaction of the substrates malonate, ATP, and CoA with malonyl-CoA synthetase, the catalytic product malonyl-CoA was characterized by NMR spectroscopy and molecular modeling. To assign proton and carbon chemical shifts, two-dimensional $^1H-^1H$ DQF-COSY and $^1H-^{13}C$ HMBC experiments were used. The structure of malonyl-CoA in the solution phase was determined based on distance constraints from NOESY and ROESY spectra. The structures were well-converged around the pantetheine region with the pairwise RMSD value of 0.08 nm. The solution structure exhibited a compact folded conformation with intramolecular hydrogen bonds among its carbonyl and hydroxyl groups. These findings will help us to understand the initial interaction of malonate and CoA with malonyl-CoA synthetase.

Keywords

Malonyl-CoA;Malonyl-CoA Synthetase;NMR;Simulated annealing

Acknowledgement

Supported by : Korea Science and Engineering Foundation