High Molecular Weight Kininogen Is Cleaved by FXIa at Three Sites: Arg⁴⁰⁹-Arg⁴¹⁰, Lys⁵⁰²-Thr⁵⁰³ and Lys³²⁵-Lys³²⁶

Summary

We investigated the cleavage of high molecular weight kininogen (HK) by activated coagulation factor XI (FXIa) in vitro. Incubation of HK with FXIa resulted in the generation of cleavage products which were subjected to SDS-Page and analyzed by silverstaining, ligandblotting and immunoblotting, respectively. Upon incubation with FXIa, bands were generated at 111, 100, 88 kDa on nonreduced and at 76, 62 and 51 kDa on reduced gels. Amino acid sequence analysis of the reaction mixtures revealed three cleavage sites at Arg⁴⁰⁹-Arg⁴¹⁰, at Lys⁵⁰²-Thr⁵⁰³ and at Lys³²⁵-Lys³²⁶. Analysis of HK-samples incubated with FXIa for 3 min, 10 min and 120 min indicated HK to be cleaved first at Arg⁴⁰⁹-Arg⁴¹⁰, followed by cleavage at Lys⁵⁰²-Thr⁵⁰³ and then at Lys³²⁵-Lys³²⁶.

In conclusion, HK is cleaved by FXIa at three sites. Cleavage of HK by FXIa results in the loss of the surface binding site of HK, which may constitute a mechanism of inactivation of HK and of control of contact system activation.

Key words

Factor XIa - high molecular weight kininogen - contact activation - cleavage site

• References

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