Immunochemical determination of conformational equilibria for fragments of the B beta chain of fibrinogen.
The conformations of the B beta chain of the intact fibrinogen molecule and of various fragments of the B beta chain of fibrinogen that contain the region that is hydrolyzed by thrombin have been compared by an immunochemical method [Sachs, D. H., Schechter, A. N., Eastlake, A., & Anfinsen, C. B. (1972) Proc. Natl. Acad. Sci. U.S.A. 69, 3790]. Anti-fibrinogen antibodies were induced in rabbits by immunization with native bovine fibrinogen. An antibody population specific for the native antigenic determinant within the B beta fragment 20-28 was isolated by immunoadsorption. This preparation was to determine the value of Kconf, the equilibrium constant for the interconversion of the nonnative and native conformations of this determinant. Values of Kconf were measured for this determinant within native fibrinogen, the disulfide knot (DSK), CNBrB beta, B beta fragment 16-28, B beta fragment 20-28, and fibrinopeptide B (FpB). 125I-Labeled fibrinogen (125I-F) was used in the determination of Kconf by measuring the competition between 125I-F and the fibrinogen derivatives under study for binding to the purified antibody. For the antigenic region in F, the DSK, and CNBrB beta, the values of Kconf at 4 degrees C were infinity, (5.9 +/- 3.5) X 10(-3), and (1.2 +/- 0.7) X 10(-3), respectively. The values of Kconf for B beta fragment 16-28, B beta fragment 20-28, and FpB at 4 degrees C were less than (6.0 +/- 3.9) X 10(-7).(ABSTRACT TRUNCATED AT 250 WORDS)
Nagy
J A
JA
Meinwald
Y C
YC
Scheraga
H A
HA
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002417:Cattle; D003488:Cyanogen Bromide; D000939:Epitopes / Q000032:analysis; D005340:Fibrinogen / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D010446:Peptide Fragments / Q000276:immunology; D011487:Protein Conformation; D013045:Species Specificity; D013917:Thrombin
D000906:Antibodies; D000939:Epitopes; D046911:Macromolecular Substances; D010446:Peptide Fragments; D005340:Fibrinogen; D013917:Thrombin; D003488:Cyanogen Bromide