Fibrinogen Mitaka II: A Hereditary Dysfibrinogen With Defective Thrombin Binding Caused by an Aα Glu-11 to Gly Substitution

Abstract

AbstractA new type of Aα Glu-11 to Gly substitution has been identified in a congenitally abnormal fibrinogen, fibrinogen Mitaka II, derived from a 14-year-old female suffering from easy bruising since childhood. Plasma of the patient and fibrinogen purified therefrom were found to clot slowly by thrombin but in a normal fashion by ancrod, a thrombin-like snake venom enzyme. The ancrod-clotted fibrin gels were normally solid and turbid, whereas the thrombin-clotted gels were initially fragile and transparent but became gradually normalized during further incubation. On reverse-phase high-performance liquid chromatography, there was an additional peptide group eluted distinctly later than the corresponding normal fibrinopeptide A in the clot-liquor of the patient’s samples. Sequence analysis of these aberrant peptides and isolated Aα chains of the patient’s fibrinogen showed that Glu at position 11 of the abnormal Aα chain had been replaced by Gly. Studies using 125l-labeled thrombin showed that the binding with thrombin was evidently reduced for her fibrinogen and the aberrant fibrinopeptide A as compared with that for the normal controls, indicating that Aα Glu-11 may be critical for the fibrinogen-thrombin interaction. Indeed, Aa Glu-11 of fibrinogen has recently been proposed to stabilize the local conformation, including the β-turn, and to form a salt bridge between its side-chain carboxyl group and the guanidino group of Arg-173 of thrombin based on crystallographic analyses using analogs of fibrinopeptide A complexed with thrombin (Stubb et al, Eur J Biochem 206:187, 1992 and Martin et al, J Biol Chem 267:7911, 1992).