Effects of carboxy-terminal truncation on human lecithin:cholesterol acyltransferase activity.
Mutagenesis was carried out in human lecithin:cholesterol acyltransferase (LCAT) to generate mutants with stop codons at positions corresponding to amino acids 315, 341, 359, 375, 388, 394, and 398 of the 416-amino acid sequence of the mature enzyme protein. Deletion of the 18 terminal amino acids of the protein was without effect on LCAT phospholipase or acyltransferase activity, or the stability of the protein to denaturation at 37 degrees C. Further deletion led to loss of most of the activity, associated with a 10-fold increase in the rate of denaturation at 37 degrees C. These data indicate that the proline-rich C-terminus of LCAT is not required for effective enzyme activity. The loss of activity that accompanied deletion of residues 394-398 suggests a structural role for these residues, part of a series of predicted beta-sheet sequences in the C-terminal third of the LCAT primary sequence.
Francone
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OL
Cardiovascular Research Institute, University of California Medical Center, San Francisco 94143, USA.
Evangelista
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L
Fielding
C J
CJ
D000818:Animals; D016632:Apolipoprotein A-I / Q000494:pharmacology; D001483:Base Sequence; D016466:CHO Cells / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism; D002788:Cholesterol Esters / Q000096:biosynthesis; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D004795:Enzyme Stability; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D010740:Phospholipases / Q000378:metabolism; D017433:Protein Structure, Secondary; D017384:Sequence Deletion
D016632:Apolipoprotein A-I; D002788:Cholesterol Esters; D018076:DNA, Complementary; D008081:Liposomes; D010713:Phosphatidylcholines; D002784:Cholesterol; D007862:Phosphatidylcholine-Sterol O-Acyltransferase; D010740:Phospholipases