Lecithin:cholesterol acyltransferase. Functional regions and a structural model of the enzyme.
The amino acid sequence of human lecithin:cholesterol acyltransferase has been determined by degradation and alignment of peptides obtained from tryptic and staphylococcal digestions and the cleavage with cyanogen bromide and consisted of 416 amino acid residues. All of the tryptic peptides of lecithin:cholesterol acyltransferase were isolated and sequenced. Peptides resulting from digestion by staphylococcal protease, cyanogen bromide cleavage, or the combination of the two methods were employed to find overlapping segments. The N terminus of human lecithin:cholesterol acyltransferase was determined to be phenylalanine by sequencing the whole protein up to 40 residues while the C terminus was identified as glutamic acid through carboxypeptidase Y cleavage. Cys50 and Cys74 and Cys313 and Cys356 were identified as the two disulfide bridges while the free sulfhydryl groups were located at positions 31 and 184. The N-glycosylated sites of the protein were assigned to asparagines at positions 20, 84, 272, and 384. The active site of lecithin:cholesterol acyltransferase was identified as serine on position 181 according to its homology with other serine-type esterases which have a common structure of glycine-variable amino acid-active serine-variable amino acid-glycine (Gly-X-Ser-X-Gly) with the variable amino acids disrupting the homology. No long internal repeats or homologies with apolipoproteins were found. The secondary structure is consistent with the results of predictive algorithms. A simple model of the enzyme is proposed on the basis of available chemical data and predictive methods.
D000595:Amino Acid Sequence; D001665:Binding Sites; D002268:Carboxypeptidases; D003488:Cyanogen Bromide; D003545:Cysteine; D004220:Disulfides; D010450:Endopeptidases; D005971:Glutamates; D018698:Glutamic Acid; D006031:Glycosylation; D006801:Humans; D010446:Peptide Fragments; D010649:Phenylalanine; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000097:blood*; D011487:Protein Conformation; D012697:Serine Endopeptidases; D014357:Trypsin
D004220:Disulfides; D005971:Glutamates; D010446:Peptide Fragments; D018698:Glutamic Acid; D010649:Phenylalanine; D007862:Phosphatidylcholine-Sterol O-Acyltransferase; D002268:Carboxypeptidases; D010450:Endopeptidases; C022633:serine carboxypeptidase; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin; D003545:Cysteine; D003488:Cyanogen Bromide