Molecular Mechanism of the Drop in the p<i>K</i><sub>a</sub> of a Substrate Analog Bound to Medium-Chain Acyl-CoA Dehydrogenase: Implications for Substrate Activation

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Author(s)

    • Nishina Yasuzo
    • Department of Molecular Physiology Graduate School of Medical Sciences, Kumamoto University
    • Sato Kyosuke
    • Department of Molecular Physiology Graduate School of Medical Sciences, Kumamoto University
    • Tamaoki Haruhiko
    • Department of Molecular Enzymology, Graduate School of Medical Sciences, Kumamoto University
    • Tanaka Takeyuki
    • Division of Biological Chemistry, Department of Life Science, Graduate School of Science and Technology, Kobe University
    • Setoyama Chiaki
    • Department of Molecular Enzymology, Graduate School of Medical Sciences, Kumamoto University
    • Miura Retsu
    • Department of Molecular Enzymology, Graduate School of Medical Sciences, Kumamoto University
    • Shiga Kiyoshi
    • Department of Molecular Physiology Graduate School of Medical Sciences, Kumamoto University

Abstract

The p<i>K</i><sub>a</sub> value of a substrate analogue 3-thiaoctanoyl-CoA at αC-H is known to drop from <i>ca</i>. 16 in the free state to 5-6 upon binding to medium-chain acyl-CoA dehydrogenase (MCAD). The molecular mechanism underlying this phenomenon was investigated by taking advantage of artificial FADs, <i>i.e</i>., 8-CN-, 7, 8-Cl<sub>2</sub>-, 8-Cl-, 8-OCH<sub>3</sub>-, 8-NH<sub>2</sub>-, ribityl-2'-deoxy-8-CN-, and ribityl-2'-deoxy-8-Cl-FADs, reconstituted into MCAD. The stronger the electron-withdrawing ability of the substituent, the smaller the p<i>K</i><sub>a</sub> value became [<i>e. g</i>., 7.4 (8-NH<sub>2</sub>-FAD) and 4.0 (8-CN-FAD)], suggesting that the flavin ring itself affects the p<i>K</i><sub>a</sub> value of the ligand <i>via</i> a charge-transfer interaction with the ligand. The destruction of the hydrogen bond between the thioester C(1)=O and the ribityl-2'-OH of FAD raised the p<i>K</i><sub>a</sub> by <i>ca</i>. 2.5 units. These results indicate that the interaction between the ligand and the flavin ring also serves to lower the p<i>K</i><sub>a</sub> of the ligand, in addition to the hydrogen bonds at C(1)=O of the ligand.

Journal

  • The Journal of Biochemistry

    The Journal of Biochemistry 134(6), 835-842, 2003

    The Japanese Biochemical Society

Codes

  • NII Article ID (NAID)
    130003534635
  • Text Lang
    ENG
  • ISSN
    0021-924X
  • Data Source
    J-STAGE 
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