Alteration of the fatty acid substrate specificity of lysophosphatidate acyltransferase by site-directed mutagenesis

Larry Zee Morand, Shilpa Patil, Mary Quasney, J. Bruce German

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

The JC201 strain of Eschericia coli contains a temperature-sensitive lesion in lysophosphatidate acyltransferase (LPAT) activity. The LPAT gene from JC201 was isolated by PCR and a single mutant nucleotide, adenine-440, was identified by DNA sequence analysis. Site-directed mutagenesis converted the mutant adenine-440 back to the native guanine-440 nucleotide. The restored LPAT gene rescued JC201 cells at the non-permissive temperature. The fatty acid substrate specificity of LPAT from Eschericia coli was altered by site-directed mutagenesis of a single amino acid in the restored LPAT gene. Threonine-122 of LPAT was changed to alanine or leucine. A change from threonine-122 to alanine increased the substrate specificity in vitro for oleoyl-CoA and linoleoyl-CoA; whereas a change to leucine increased the substrate specificity for lignoceroyl-CoA.

Original languageEnglish (US)
Pages (from-to)79-84
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume244
Issue number1
DOIs
StatePublished - Mar 6 1998

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Fingerprint Dive into the research topics of 'Alteration of the fatty acid substrate specificity of lysophosphatidate acyltransferase by site-directed mutagenesis'. Together they form a unique fingerprint.

  • Cite this