Undecaprenyl pyrophosphate synthetase from Lactobacillus plantarum: A dimeric protein

https://doi.org/10.1016/0003-9861(84)90085-7Get rights and content

Abstract

Undecaprenyl pyrophosphate synthetase has been purified from Lactobacillus plantarum. It catalyzes the formation of a C55 polyprenyl pyrophosphate having isoprene residues with cis stereochemistry. The enzyme was shown to be an acidic protein (pI = 5.1), which can be partially purified by preparative gel electrophoresis and Blue-agarose column chromatography. The Km's of the enzyme for its substrates t,t-farnesyl pyrophosphate and isopentenyl pyrophosphate were determined to be 0.13 and 1.92 μm, respectively. The molecular weight of the enzyme was estimated by molecular sieve chromatography and gradient centrifugation to be 56,000 ± 4000. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the protein was composed of a dimer of 30,000-Da subunits. The enzyme was inactivated by the arginine-specific reagents phenylglyoxal, butanedione and, cyclohexanedione, but this inactivation was not prevented by either of the substrates.

References (42)

  • J.G. Christenson et al.

    J. Biol. Chem

    (1969)
  • T. Kurokawa et al.

    Biochem. Biophys. Res. Commun

    (1971)
  • T. Baba et al.

    Arch. Biochem. Biophys

    (1980)
  • M.V. Keenan et al.

    Arch. Biochem. Biophys

    (1974)
  • C.M. Allen et al.

    Arch. Biochem. Biophys

    (1976)
  • D.K. Grange et al.

    Biochem. Biophys. Res. Commun

    (1977)
  • R.B. Wellner et al.

    FEBS Lett

    (1979)
  • G.F. Barnard et al.

    Biochim. Biophys. Acta

    (1980)
  • J.R. Kalin et al.

    Biochim. Biophys. Acta

    (1979)
  • O.H. Lowry et al.

    J. Biol. Chem

    (1951)
  • L. Shuster
  • K. Weber et al.
  • C.D. Poulter et al.

    J. Biol. Chem

    (1978)
  • H. Fujii et al.

    J. Biol. Chem

    (1982)
  • H. Jäckle

    Anal. Biochem

    (1979)
  • W. Wray et al.

    Anal Biochem

    (1981)
  • J.D. Muth et al.

    Biochim. Biophys. Acta

    (1979)
  • L. Weng et al.

    J. Biol. Chem

    (1978)
  • T.K. Wong et al.

    J. Biol. Chem

    (1982)
  • W.L. Adair et al.

    J. Biol. Chem

    (1982)
  • K. Momose et al.

    J. Biol. Chem

    (1972)
  • Cited by (21)

    • Activation and inhibition of rubber transferases by metal cofactors and pyrophosphate substrates

      2003, Phytochemistry
      Citation Excerpt :

      In addition, the study of biochemical parameters on the native particles is likely to provide kinetic information directly applicable to the in vivo situation, something always questionable in the study of solubilized membrane-bound enzymes. Other cis-prenyl transferases have been studied: the soluble, prokaryotic undecaprenyl pyrophosphate synthase (UPPS) (Baba and Allen, 1978; Muth and Allen, 1984; Shimizu et al., 1998; Apfel et al., 1999; Hemmi et al., 2001; Kharel et al., 2001; Ko et al., 2001) and the membrane-associated, eucaryotic dehydrodolichyl pyrophosphate synthase (DDPPS) (Adair and Cafmeyer, 1987; Matsuoka et al., 1991; Ericsson et al., 1992; Sato et al., 1999; Chang et al., 2001). Unlike RuT, both UPPS and DDPPS have products with defined molecular weights.

    View all citing articles on Scopus

    This work was supported by USPHS Grant GM 23193.

    View full text