Cysteine188 Revealed as Being Critical for the Enzyme Activity of Arylmalonate Decarboxylase by Site-Directed Mutagenesis

Access this Article

Search this Article

Author(s)

Abstract

Arylmalonate decarboxylase (AMDase) catalyzes the asymmetric decarboxylation of <i>α</i>-aryl-<i>α</i>-methylmalonic acid. Since this enzyme is inhibited by SH-reagents, a cysteine residue is supposed to be involved at the catalytic site. Cloning of the gene which codes the enzyme revealed that this enzyme contains four cysteine residues. Titration of free SH groups by <i>p</i>-(chloromercurio)benzoate disclosed that all four Cys are in the reduced form. In this study, four mutant enzymes (C101S, C148S, C171S, and C188S) were prepared, in which one of four cysteines was replaced by serine. The CD spectra indicated that the conformational differences of C101S and C188S compared to that of the native enzyme are not so significant. The catalytic activities of the four mutants were measured. Among these mutant enzymes, only C188S showed a drastic decrease in enzyme activity, indicating that cysteine<sup>188</sup> is located at the active center of the enzyme. The catalytic activities of the other mutants are also discussed.

Journal

  • Bulletin of the Chemical Society of Japan

    Bulletin of the Chemical Society of Japan 70(11), 2765-2769, 1997-11-15

    The Chemical Society of Japan

References:  13

  • <no title>

    MIYAMOTO K.

    J. Am. Chem. Soc. 112, 4077, 1990

    Cited by (10)

  • <no title>

    MIYAMOTO K.

    Biocatalysis 5, 49, 1990

    Cited by (1)

  • <no title>

    MIYAMOTO K.

    J. Fluorine Chem. 59, 225, 1992

    Cited by (2)

  • <no title>

    MIYAMOTO K.

    Eur. J. Biochem. 210, 475, 1992

    Cited by (3)

  • <no title>

    MIYAMOTO K.

    Appl. Microbiol Biotechnol. 38, 234, 1992

    Cited by (2)

  • <no title>

    KAWASAKI T.

    Bull. Chem. Soc. Jpn. 68, 2017, 1995

    Cited by (6)

  • <no title>

    KAWASAKI T.

    Chem. Lett. 1996, 195

    Cited by (2)

  • <no title>

    KUNKEL T. A.

    Methods Enzymol. 154, 367, 1987

    Cited by (3)

  • <no title>

    BAHATTACHARYYA D. K.

    J. Biol. Chem. 271, 2179, 1996

    Cited by (2)

  • <no title>

    HASHIMOTO Y.

    J. Biochem. 119, 145, 1996

    Cited by (2)

  • <no title>

    MIYAMOTO K.

    Bioorg. Med. Chem. 2, 469, 1994

    Cited by (5)

  • <no title>

    KAWASAKI T.

    Bull.Chem.Soc.Jpn. 69, 3591, 1996

    Cited by (3)

  • <no title>

    MOHAMEDALI K.

    Biochemistry 35, 1672, 1996

    DOI  Cited by (2)

Cited by:  1

Codes

  • NII Article ID (NAID)
    10008925465
  • NII NACSIS-CAT ID (NCID)
    AA00580132
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    00092673
  • NDL Article ID
    4346210
  • NDL Source Classification
    ZP1(科学技術--化学・化学工業)
  • NDL Call No.
    Z53-B35
  • Data Source
    CJP  CJPref  NDL  J-STAGE 
Page Top