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 language | English (US) |
|---|---|
| Pages (from-to) | 79-84 |
| Number of pages | 6 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 244 |
| Issue number | 1 |
| DOIs | |
| State | Published - Mar 6 1998 |
Fingerprint
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 journal › Article
}
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 -