Self-association of the major protein component of bovine serum high density lipoprotein.
The major protein component of bovine high density lipoprotein was investigated in solution by fluorescence polarization and ultracentrifugal techniques. A fluorescent derivative of this protein with 1-dimethylaminonaphthalene-5-sulfonyl chloride was employed in the fluorescence experiments. Over the concentration range from 5-10(-7) M to 5-10(-4) M of the protein monomer at pH values from 2 to 11 and ionic strengths from 0.03 to 2.0, at 23 degrees C, the major protein of bovine high density lipoproteinapoprotein (Apo-HOL-I) was found to exist in a stable aggregated form. The aggregate was not affected by dioxane additions of up to 20% nor by Triton X-100 to 0.2%, but dissociated readily in the presence of 0.07% sodium dodecylsulfate or 6 M urea. At concentrations below 5-10(-7) M, dissociation of the protein aggregate started spontaneously and continued down to 10(-8) M, the lowest measurable concentration. Several physiocochemical properties of the major protein of bovine high density lipoprotein were determined in the stable aggregate form. Molecular weight was 104 000 from ultracentrifugal analysis and 80 000 from gel-filtration. Rotational relaxation time was 115 ns at 25 degrees C, and s-0 20,w was 4.78 s. The results suggest very strong protein-protein interactions (Kd less than 10(-7) M) that are not electrostatic in nature. Hydrophobic interactions of a magnitude that could be affected by 20% dioxane or 0.2% Triton X-100 detergent are also excluded. There is saturation of the interaction sites by the aggregation of a few protein monomer units possibly to form a tetramer which is moderately asymmetric (1:4 axial ratio, assuming an ellipsoid of revolution) and relatively rigid. The strong protein-protein interactions in this pure apolipoprotein suggest the possibility of competition of inter-protein associations with protein-lipid interactions in in vitro lipid binding or lipoprotein reconstitution experiments.
D000818:Animals; D001059:Apoproteins / Q000097:blood*; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003619:Dansyl Compounds; D004146:Dioxanes; D005453:Fluorescence; D008075:Lipoproteins, HDL / Q000097:blood*; D008970:Molecular Weight; D009904:Optical Rotation; D011092:Polyethylene Glycols / Q000494:pharmacology; D011487:Protein Conformation; D012967:Sodium Dodecyl Sulfate; D013696:Temperature; D014461:Ultracentrifugation; D014508:Urea
D001059:Apoproteins; D003619:Dansyl Compounds; D004146:Dioxanes; D008075:Lipoproteins, HDL; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D014508:Urea