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 Citations (Scopus)

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

Fingerprint

Acyltransferases
Mutagenesis
Substrate Specificity
Site-Directed Mutagenesis
Fatty Acids
Substrates
Genes
Threonine
Leucine
Alanine
Temperature
Guanine Nucleotides
Adenine Nucleotides
DNA sequences
Guanine
Adenine
DNA Sequence Analysis
Nucleotides
Amino Acids
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Alteration of the fatty acid substrate specificity of lysophosphatidate acyltransferase by site-directed mutagenesis. / Morand, Larry Zee; Patil, Shilpa; Quasney, Mary; German, J. Bruce.

In: Biochemical and Biophysical Research Communications, Vol. 244, No. 1, 06.03.1998, p. 79-84.

Research output: Contribution to journalArticle

Morand, Larry Zee ; Patil, Shilpa ; Quasney, Mary ; German, J. Bruce. / Alteration of the fatty acid substrate specificity of lysophosphatidate acyltransferase by site-directed mutagenesis. In: Biochemical and Biophysical Research Communications. 1998 ; Vol. 244, No. 1. pp. 79-84.
@article{af36f7d6f4954399b80ca37009574718,
title = "Alteration of the fatty acid substrate specificity of lysophosphatidate acyltransferase by site-directed mutagenesis",
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.",
author = "Morand, {Larry Zee} and Shilpa Patil and Mary Quasney and German, {J. Bruce}",
year = "1998",
month = "3",
day = "6",
doi = "10.1006/bbrc.1998.8218",
language = "English (US)",
volume = "244",
pages = "79--84",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Alteration of the fatty acid substrate specificity of lysophosphatidate acyltransferase by site-directed mutagenesis

AU - Morand, Larry Zee

AU - Patil, Shilpa

AU - Quasney, Mary

AU - German, J. Bruce

PY - 1998/3/6

Y1 - 1998/3/6

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0032489279&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032489279&partnerID=8YFLogxK

U2 - 10.1006/bbrc.1998.8218

DO - 10.1006/bbrc.1998.8218

M3 - Article

C2 - 9514885

AN - SCOPUS:0032489279

VL - 244

SP - 79

EP - 84

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -