Nucleotide sequence of the cDNA for lecithin-cholesterol acyl transferase (LCAT) from the rat

Abstract

We report here the nucleotide and deduced amino acid sequence for the lecithin-cholesterol acyl transferase (LCAT) gene from the rat. Screening a X GT 10 cDNA hepatic library with a partial human LCAT cDNA as a probe (1) resulted in two overlapping cDNA clones whose sequence contains an open reading frame that codes for a 440 amino acid polypeptide plus short regions of 5' and 3' untranslated sequences.Sequence comparison of the cDNA deduced polypeptide derived from the rat and human revealed 86.4% of homology while at the nucleotide level the degree of sequence homology is 84.5%. Likewise rat-mouse sequences are 91.6% and 89.6% homologous at the amino acid and nucleotide levels respectively.All the main structural features already described for the human and mouse protein sequences were also present in the rat LCAT enzyme (2-3) and in particular an amino acid Arg 147 of the human protein whose substitution to Trp (CGG-TGG) was found to be responsible for LCAT enzyme inactivation in a patient homozygous for LCAT deficiency is conserved in the rat protein as well (4). ACKNOWLEDGEMENTS

Quantitative trait loci influencing cholesterol and phospholipid phenotypes map to chromosomes that contain genes regulating blood pressure in the spontaneously hypertensive rat.

“…The Lcat gene, coding for lecithin cholesterol acyl transferase, has been mapped by finding an RFLP of the PCR products between SHR and BN.Lx strains. The primers were designed according to the published cDNA sequence (25) to amplify the last intron of the Lcat gene as based on homology with the human complete DNA sequence (26): the upstream primer was 5Ј-TGA TGG CTT CAT CTC TCT CG, the downstream primer was 5Ј-AGG GGG AAG TTG TGG TTA TG. The size of the uncut PCR product was ‫ف‬ 1,350 bp.…”

Synthesis and secretion of wild-type and mutant human plasma cholesteryl ester transfer protein in baculovirus-transfected insect cells: the carboxyl-terminal region is required for both lipoprotein binding and catalysis of transfer.

“…As shown in Table 3, there is a considerable sequence similarity between octapeptides in CETP (7,34), lecithin:cholesterol acyltransferase (LCAT) (29,35,36), pancreatic cholesterol esterase (30), bactericidal permeability increasing protein (33), lipopolysaccharide binding protein (24), cholesterol 7a-hydroxylase (32), and hormone-sensitive lipase (33). The similarity is also marked in the corresponding nucleotide sequences.…”

Amino Acid Residue 149 of Lecithin:Cholesterol Acyltransferase Determines Phospholipase A2 and Transacylase Fatty Acyl Specificity

“…Subsequent studies with purified enzyme showed that rat LCAT was more reactive with PC substrates containing arachidonic acid compared to human LCAT (16 -19). This difference occurs in spite of a 87% identity between the rat and human LCAT protein sequence (20,21). The molecular explanation for the difference in reactivity is * This work was supported by National Institutes of Health Grants HL 49373 (to J. S. P.), HL 54176 (to J. S. P.), and HL 47864 (to R. A.…”