Novel N-terminal mutation of human apolipoprotein A-I reduces self-association and impairs LCAT activation.
We have identified a novel mutation in apoA-I (serine 36 to alanine; S36A) in a human subject with severe hypoalphalipoproteinemia. The mutation is located in the N-terminal region of the protein, which has been implicated in several functions, including lipid binding and lecithin:cholesterol acyltransferase (LCAT) activity. In the present study, the S36A protein was produced recombinantly and characterized both structurally and functionally. While the helical content of the mutant protein was lower compared with wild-type (WT) apoA-I, it retained its helical character. The protein stability, measured as the resistance to guanidine-induced denaturation, decreased significantly. Interestingly, native gel electrophoresis, cross-linking, and sedimentation equilibrium analysis showed that the S36A mutant was primarily present as a monomer, notably different from the WT protein, which showed considerable oligomeric forms. Although the ability of S36A apoA-I to solubilize phosphatidylcholine vesicles and bind to lipoprotein surfaces was not altered, a significantly impaired LCAT activation compared with the WT protein was observed. These results implicate a region around S36 in apoA-I self-association, independent of the intact C terminus. Furthermore, the region around S36 in the N-terminus of human apoA-I is necessary for LCAT activation.
Weers
Paul M M
PM
Department of Chemistry and Biochemistry, California State University Long Beach, Long Beach, CA, USA.
Patel
Arti B
AB
Wan
Leon C-P
LC
Guigard
Emmanuel
E
Kay
Cyril M
CM
Hafiane
Anouar
A
McPherson
Ruth
R
Marcel
Yves L
YL
Kiss
Robert S
RS
D016632:Apolipoprotein A-I / Q000737:chemistry* / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D009154:Mutation; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000737:chemistry / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D013329:Structure-Activity Relationship
D016632:Apolipoprotein A-I; D010713:Phosphatidylcholines; D007862:Phosphatidylcholine-Sterol O-Acyltransferase