Rat sn-glycerol-3-phosphate acyltransferase: molecular cloning and characterization of the cDNA and expressed protein

• Published in: Biochimica et biophysica acta 1999-08, Vol.1439 (3), p.415-423
• Main Authors: Ganesh Bhat, B, Wang, Ping, Kim, Ji-Hyeon, Black, Tracy M, Lewin, Tal M, Fiedorek, Frederick T, Coleman, Rosalind A
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cell Line; Cloning, Molecular; Diacylglycerol metabolism; DNA, Complementary - chemistry; DNA, Complementary - genetics; Glycerol-3-phosphate acyltransferase; Glycerol-3-Phosphate O-Acyltransferase - biosynthesis; Glycerol-3-Phosphate O-Acyltransferase - chemistry; Glycerol-3-Phosphate O-Acyltransferase - genetics; Glycerolipid synthesis; Insecta; Mitochondria, Liver - enzymology; Molecular Sequence Data; Rats; Sequence Alignment; Triacylglycerol
• ISSN: 1388-1981; 0006-3002; 1879-2618
• DOI: 10.1016/S1388-1981(99)00103-1

Rat mitochondrial glycerol-3-phosphate acyltransferase (GPAT) cDNA was cloned and characterized. We identified a cDNA containing an open reading frame of 828 amino acids that had an 89% homology with the coding region of the previously characterized mouse mitochondrial GPAT and a predicted amino acid sequence that was 96% identical. The rat 5′ UTR was only 159 nucleotides, in contrast to the 926 nucleotide 5′ UTR of the mouse cDNA and had an internal deletion of 167 nucleotides. GPAT was expressed in Sf21 insect cells, and specific inhibitors strongly suggest that, like the Escherichia coli GPAT, the recombinant mitochondrial GPAT and the mitochondrial GPAT isoform in rat liver contain critical serine, histidine, and arginine residues.