Protein lipid interaction in bile: effects of biliary proteins on the stability of cholesterol-lecithin vesicles.
The nucleation of cholesterol crystals is an obligatory precursor to cholesterol gallstone formation. Nucleation, in turn, is believed to be preceded by aggregation and fusion of cholesterol-rich vesicles. We have investigated the effects of two putative pro-nucleating proteins, a concanavalin A-binding protein fraction and a calcium-binding protein, on the stability of sonicated small unilamellar cholesterol-lecithin vesicles. Vesicle aggregation is followed by monitoring absorbance, and upon addition of the concanavalin A-binding protein fraction the absorbance of a vesicle dispersion increases continuously with time. Vesicle fusion is probed by a fluorescence contents-mixing assay. Vesicles apparently fuse slowly after the addition of the concanavalin A-binding protein, although inner filter effects confound the quantitative measurement of fusion rates. The rates of change of absorbance and fluorescence increase with the concentration of the protein, and the second-order dimerization rate constant increases with both the protein concentration and the cholesterol content of the vesicles. On the other hand, the calcium-binding protein has no effect on the stability of the vesicle dispersion. This protein may therefore affect cholesterol crystal formation not by promoting the nucleation process, but by enhancing crystal growth and packaging. Our results demonstrate that biliary proteins can destabilize lipid vesicles and that different proteins play different roles in the mechanism of cholesterol gallstone formation.
Luk
A S
AS
Department of Chemical Engineering, Center for Molecular and Engineering Thermodynamics, University of Delaware, Newark, DE, USA.
Kaler
E W
EW
Lee
S P
SP
D001646:Bile / Q000378:metabolism*; D002769:Cholelithiasis / Q000209:etiology; D002784:Cholesterol / Q000737:chemistry*; D003460:Crystallization; D019281:Dimerization; D006801:Humans; D050356:Lipid Metabolism; D008956:Models, Chemical; D010713:Phosphatidylcholines / Q000737:chemistry*; D011506:Proteins / Q000378:metabolism*; D011952:Receptors, Concanavalin A / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
D010713:Phosphatidylcholines; D011506:Proteins; D011952:Receptors, Concanavalin A; C027774:concanavalin A-binding glycoproteins; D002784:Cholesterol