Conformation of the carboxy-terminal region of the A alpha chain of fibrinogen as elucidated by immunochemical analyses.
The conformation of the carboxy-terminal aspects of the A alpha chain of human fibrinogen has been assessed by immunochemically characterizing the A alpha 239-476 and A alpha 518-584 regions of the molecule. Two peptides, corresponding to these regions, were isolated from cyanogen bromide digests of the A alpha chain by molecular exclusion and high-performance liquid chromatography. Each peptide reacted with antibodies elicited by immunization with the A alpha chain and intact fibrinogen. A alpha 239-476 appears to be a relatively immunodominant region of the molecule. Competitive inhibition analyses confirmed the accessibility of these regions to antibody in native fibrinogen. Each peptide, however, contained one or more epitopes, which was occult in the native molecule. These occult epitopes were expressed by the intact A alpha chain and became accessible when fibrinogen was cleaved with plasmin. With plasmic degradation the epitopes expressed by fibrinogen and contained within these two peptide regions became significantly more reactive with antibody. This change occurred in concert with release of the A alpha 518-584 region from the core of the molecule but did not require the generation of free A alpha 239-476. Ultimately the epitopes within both regions were shed from the plasmin-resistant core of fibrinogen. Peptide epitopes were expressed in a similar manner by prolonged plasmic degradation of fibrinogen and fibrin with alpha chain cross-linking. These results are generally consistent with models depicting the carboxy-terminal aspects of the A alpha chain as being surface-oriented but suggest a systematic ordering of structure when these regions are integrated into the native molecule. Plasmic cleavage significantly relaxes the conformational restraints on the organization within this region.
D000596:Amino Acids / Q000032:analysis; D001667:Binding, Competitive; D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D005340:Fibrinogen / Q000276:immunology; D006801:Humans; D010446:Peptide Fragments / Q000276:immunology; D011487:Protein Conformation; D011863:Radioimmunoassay / Q000379:methods
D000596:Amino Acids; D000939:Epitopes; D010446:Peptide Fragments; D005340:Fibrinogen