Binding of 25-hydroxycholesterol and cholesterol to different cytoplasmic proteins.
Studies were carried out to determine whether or not oxygenated derivatives of cholesterol (e.g., 25-hydroxycholesterol) that specifically suppress the activity of 3-hydroxy-3-methylglutaryl-CoA reductase [mevalonate:NADP(+) oxidoreductase (CoA-acylating), EC 1.1.1.34], bind to a soluble component of the cytoplasm different from that which binds the nonsuppressor, cholesterol. Density gradient fractionation of the cytosolic fraction isolated from L cell cultures that had been incubated with low concentrations of 25-hydroxy[26,27-(3)H]cholesterol or [1,2-(3)H]cholesterol provided evidence for the existence of at least two different sterol-binding proteins. Bound cholesterol sedimented in a sucrose density gradient as two or more broad bands with coefficients of approximately 9 S and 21 S. Two relatively narrow bands of bound 25-hydroxycholesterol had sedimentation coefficients of 5 S and 8 S. Preincubation of the cells with a relatively high concentration of unlabeled 25-hydroxycholesterol altered the banding pattern of the 25-hydroxy[(3)H]cholesterol taken up during a subsequent incubation period by decreasing the size of the major (8S) band. Under these conditions, cholesterol did not affect the banding pattern of 25-hydroxy[(3)H]cholesterol. The density gradient banding pattern of bound [(3)H]cholesterol was only slightly affected by preincubating the cells with unlabeled cholesterol or 25-hydroxycholesterol. Both sterols appeared to be bound to proteins because the bound sterols were eliminated from cytosol that had been heated at 100 degrees , and their sedimentation coefficients were altered by proteolysis.
D001667:Binding, Competitive; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D006888:Hydroxycholesterols / Q000378:metabolism*; D007739:L Cells / Q000378:metabolism
D002352:Carrier Proteins; D006888:Hydroxycholesterols; D002784:Cholesterol
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