Fibrinogen-sepharose interaction with prothrombin, prethrombin 1, prethrombin 2 and thrombin.
Binding of prothrombin, prethrombin 1, prethrombin 2 and thrombin to fibrinogen-Sepharose was studied. Thrombin and prethrombin 2 bound to fibrinogen-Sepharose, while prethrombin 1 and prothrombin did not. Bound thrombin and prethrombin 2 were recovered from the column by eluting with 0.1 M NaCl/0.05 M Tris-HCl buffer (pH 7.4). The affinity of thrombin and prethrombin 2 to fibrinogen-Sepharose depended on ionic strength and reached a maximum at 50 mm concentration. Prethrombin 2 interacts with fibrinogen as well as thrombin; and prothrombin fragment 1.2 is not important in the formation of this complex. Thus, prethrombin 2, which is a precursor of thrombin without measurable enzymatic activity and which lacks the single cleavage at Arg-322-Ile-323 present in thrombin, has the same or very similar structural conformation as thrombin and has the same macromolecular substrate recognition site. These results confirm the earlier results that active center is not necessary in fibrinogen-thrombin interaction.
D000595:Amino Acid Sequence; D002846:Chromatography, Affinity / Q000379:methods; D004792:Enzyme Precursors / Q000302:isolation & purification / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D008970:Molecular Weight; D011516:Prothrombin / Q000302:isolation & purification / Q000378:metabolism*; D012685:Sepharose / Q000378:metabolism*; D013917:Thrombin / Q000302:isolation & purification / Q000378:metabolism*
D004792:Enzyme Precursors; D011516:Prothrombin; D005340:Fibrinogen; D012685:Sepharose; C012007:prethrombins; D013917:Thrombin