Localization of the thrombin-binding domain on prothrombin fragment 2.

The Journal of Biological Chemistry
P C LiawJeffrey I Weitz

Abstract

Co-crystallographic studies have shown that the interaction of human prothrombin fragment 2 (F2) with thrombin involves the formation of salt bridges between the kringle inner loop of F2 and anion-binding exosite II of thrombin. When F2 binds to thrombin, it has been shown to evoke conformational changes at the active site and at exosite I of the enzyme. Using plasma, recombinant, and synthetic F2 peptides (F2, rF2, and sF2, respectively) we have further localized the thrombin-binding domain on F2. F2, rF2-(1-116), rF2-(55-116), and sF2-(63-116), all of which contain the kringle inner loop (residues 64-93) and the acidic COOH-terminal connecting peptide (residues 94-116), bind to thrombin-agarose. In contrast, analogues of the kringle inner loop, sF2-(63-90), or the COOH-terminal connecting peptide, sF2-(92-116), do not bind. Thus, contrary to predictions from the crystal structure, the COOH-terminal connecting peptide as well as the kringle inner loop are involved in the interaction of F2 with thrombin. F2 and sF2-(63-116) bind saturably to fluorescently labeled active site-blocked thrombin with Kd values of 4.1 and 51.3 microM, respectively. The affinity of sF2-(63-116) for thrombin increases about 5-fold (Kd = 10 microM) whe...Continue Reading

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Citations

Mar 6, 2007·Journal of Thrombosis and Thrombolysis·Swapan Kumar Dasgupta, Perumal Thiagarajan
Jun 20, 2003·Progress in Lipid Research·Barry R Lentz
Jul 11, 2009·The Journal of Biological Chemistry·Nicolas S PetreraJeffrey I Weitz
Jun 19, 2008·Biochimica Et Biophysica Acta·Fábio R MeloPaulo A S Mourão
Oct 4, 2007·British Journal of Haematology·Rehana S LovelyDavid H Farrell
Mar 3, 2005·Biosensors & Bioelectronics·T M A GronewoldM Famulok
Dec 2, 2004·Biophysical Chemistry·James C FredenburghJeffrey I Weitz
Jun 19, 2004·The International Journal of Biochemistry & Cell Biology·Robson Q MonteiroM Lucia Bianconi
Jul 18, 2006·The International Journal of Biochemistry & Cell Biology·Helena C CastroRussolina B Zingali
Nov 1, 2016·Journal of Biomolecular Structure & Dynamics·Jiajie Xiao, Freddie R Salsbury
Feb 5, 2003·The Journal of Biological Chemistry·Lev BeckerMarlys L Koschinsky
Apr 5, 2000·The Journal of Biological Chemistry·P J AndersonP E Bock
Nov 29, 2001·The Journal of Biological Chemistry·Ingrid M VerhammePaul E Bock
Apr 19, 2003·The Journal of Biological Chemistry·Caroline H PospisilJeffrey I Weitz
Jan 8, 2000·The Journal of Biological Chemistry·P E MarqueB F Le Bonniec

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