Self-association and phospholipid binding properties of iodinated apolipoprotein A-I.
Kinetic turnover studies of apolipoprotein metabolism often utilize radioiodinated tracers. These studies rely on the "tracer assumption" that the modified tracer is physiologically and metabolically identical with the native unmodified tracer. This paper addresses the validity of this assumption on the basis of the examination of the state of self-association and binding properties with egg yolk phosphatidylcholine small unilamellar vesicles of native and iodinated apolipoprotein A-I (apoA-I). Human apoA-I was iodinated to the extent of 1.0 and 3.7 mol of nonradioactive iodine/mol of protein. At concentrations from 0.013 to 0.8 mg/mL, iodinated apoA-I underwent concentration-dependent self-association similar to that of native apoA-I as evidenced by circular dichroism and gel filtration. At all concentrations, however, the iodinated preparations were more highly self-associated as judged by gel filtration in relation to the extent of iodination. Scatchard analysis of fluorometric titrations of apoA-I/vesicle interactions demonstrated that the binding capacity of vesicles for apoA-I increased and apoA-I binding affinity decreased upon iodination. In addition, the kinetics of apoA-I binding to vesicles was enhanced by iodination. The affinity, capacity, and kinetics of apoA-I binding were each altered 2-3-fold dependent on the extent of iodination. Since the dynamic interactions of apoA-I are perturbed by iodination, one may legitimately question whether the "tracer assumption" is valid for 125I-apoA-I under all experimental conditions.
D016632:Apolipoprotein A-I; D001054:Apolipoproteins A / Q000097:blood*; D004530:Egg Yolk; D006801:Humans; D007457:Iodine Radioisotopes; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000097:blood*; D046911:Macromolecular Substances; D010713:Phosphatidylcholines / Q000378:metabolism*; D011485:Protein Binding; D013050:Spectrometry, Fluorescence
D016632:Apolipoprotein A-I; D001054:Apolipoproteins A; D007457:Iodine Radioisotopes; D008075:Lipoproteins, HDL; D046911:Macromolecular Substances; D010713:Phosphatidylcholines