V34I and V34A substitutions within the factor XIII activation peptide segment (28-41) affect interactions with the thrombin active site

Toni A. Trumbo; Muriel C. Maurer

doi:10.1055/s-0037-1613570

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2003; 89(04): 647-653
DOI: 10.1055/s-0037-1613570

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

V34I and V34A substitutions within the factor XIII activation peptide segment (28-41) affect interactions with the thrombin active site

Toni A. Trumbo

¹Department of Chemistry, University of Louisville, Louisville, Kentucky, U.S.A.

,

Muriel C. Maurer

¹Department of Chemistry, University of Louisville, Louisville, Kentucky, U.S.A.

› Institutsangaben

Abstract

Summary

In the blood coagulation cascade, thrombin helps activate Factor XIII by cleaving the Factor XIII Activation Peptide at the R37-G38 peptide bond. The common polymorphism V34L yields a Factor XIII that is more easily activated than the wild-type enzyme. Peptides based on the Factor XIII (28-41) ( ²⁸TVELQGVVPRGVNL⁴¹) sequence serve as an important model system for evaluating how to regulate thrombin activation of Factor XIII and subsequently fibrin clot character. Kinetic and NMR (1D proton line broadening and 2D transferred NOESY) studies have revealed that the P₄-P₁ region of the activation peptides is critical for binding to the thrombin surface. These results have led to an interest in exploring two new mutations at the P₄ position including V34I and V34A. The V34I peptide was found to have the lowest K_m of the peptides studied. However, unlike the V34L peptide, there are no P₄ to P₂ interactions observed in the transferred NOESY spectrum. The Leu thus promotes a bound conformation that cannot be achieved with the similar amino acid Ile. The V34A peptide exhibited a decrease in K_m but also a substantial decrease in k_cat. With this smaller amino acid, 1D proton line broadening NMR studies indicate that further contact of the Q32 residue with the thrombin surface is now possible. From these studies, valuable kinetic and structural information is being obtained to characterize interactions between thrombin and the Factor XIII activation peptide.

Theme paper: Part of this paper was originally presented at the joint meetings of the 16th International Congress of the International Society of Fibrinolysis and Proteolysis (ISFP) and the 17th International Fibrinogen Workshop of the International Fibrinogen Research Society (IFRS) held in Munich, Germany, September, 2002.

Keywords

Factor XIII - thrombin - V34L polymorphism - kinetics - transferred NOESY

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Referenzen

References
1 Mosesson MW, Siebenlist KR, Meh DA. The structure and biological features of fibrino-gen and fibrin. Ann NY Acad Sci 2001; 936: 11-30.
2 Ariens RAS, Lai T-S, Weisel JW, Greenberg CS, Grant PJ. Role of factor XIII in fibrin clot formation and effects of genetic polymorphisms. Blood 2002; 100: 743-54.
3 Ariens RAS, Philippou H, Nagaswami C, Weisel JW, Lane DA, Grant PJ. The factor XIII V34L polymorphism accelerates thrombin activation of factor XIII and affects cross-linked fibrin structure. Blood 2000; 96: 988-95.
4 Trumbo TA, Maurer MC. Examining thrombin hydrolysis of the Factor XIII Activation Peptide segment leads to a proposal for explaining the cardioprotective effects observed with the Factor XIII V34L Mutation. J Biol Chem 2000; 275: 20627-31.
5 Trumbo TA, Maurer MC. Thrombin hydrolysis of V29F and V34L mutants of Factor XIII (28-41) reveals roles of the P₉and P₄positions in Factor XIII activation. Biochemistry 2002; 41: 2859-68.
6 Sandasivan C, Yee VC. Interaction of the factor XIII activation peptide with alpha -throm-bin. Crystal structure of its enzyme-substrate analog complex. J Biol Chem 2000; 275: 36942-8.
7 Ichinose A, Davie EW. Characterization of the gene for the a subunit of human factor XIII (plasma transglutaminase), a blood coagulation factor. Proc Natl Acad Sci U.S.A 1988; 85: 5829-33.
8 Marsh Jr. HC, Meinwald YC, Thannhauser TW, Scheraga HA. Mechanism of action of thrombin on fibrinogen. Kinetic evidence for involvement of aspartic acid at position P10. Biochemistry 1983; 22: 4170-4.
9 Marsh Jr. HC, Meinwald YC, Lee S, Scheraga HA. Mechanism of action of thrombin on fib-rinogen. Direct evidence for the involvement of phenylalanine at position P9. Biochemistry 1982; 21: 6167-71.
10 Scheraga HA. Interaction of thrombin and fibrinogen and the polymerization of fibrin monomer. Ann N Y Acad Sci 1983; 408: 330-43.
11 Martin PD, Robertson W, Turk D, Huber R, Bode W, Edwards BFP. The structure of residues 7-16 of the A alpha-chain of human fibrinogen bound to bovine thrombin at 2.3-Å resolution. J Biol Chem 1992; 267: 7911-20.
12 Ni F, Meinwald YC, Vásquez M, Scheraga H A. High-resolution NMR studies of fibrinogen-like peptides in solution: structure of a thrombin-bound peptide corresponding to residues 7-16 of the A alpha chain of human fibrinogen. Biochemistry 1989; 28: 3094-105.
13 Ni F, Zhu Y, Scheraga HA. Thrombin-bound structures of designed analogs of human fib-rinopeptide A determined by quantitative transferred NOE spectroscopy: a new structural basis for thrombin specificity. J Mol Biol 1995; 252: 656-71.
14 Ni F, Ripoll DR, Martin PD, Edwards BFP. Solution structure of a platelet receptor peptide bound to bovine alpha-thrombin. Biochemistry 1992; 31: 11551-7.
15 Maurer MC, Peng J-L, An SS, Trosset J-Y, Henschen-Edman A, Scheraga HA. Structural examination of the influence of phosphorylation on the binding of fibrinopeptide A to bovine thrombin. jBiochemistry 1998; 37: 5888-902.
16 Maurer MC, Trosset JY, Lester CC, DiBella EE, Scheraga HA. New general approach for determining the solution structure of a ligand bound weakly to a receptor: structure of a fib-rinogen Aalpha-like peptide bound to throm-bin (S195A) obtained using NOE distance constraints and an ECEPP/3 flexible docking program. Proteins 1999; 34: 29-48.
17 Bode W, Turk D, Karshikov A. The refined 1.9-Å X-ray crystal structure of D-Phe-Pro-Arg chloromethylketone-inhibited human alpha-thrombin: structure analysis, overall structure, electrostatic properties, detailed active-site geometry, and structure-function relationships. Protein Sci 1992; 1: 426-79.
18 Dang QD, Sabetta M, Di Cera E. Selective loss of fibrinogen clotting in a loop-less thrombin. J Biol Chem 1997; 272: 19649-51.
19 Stubbs MT, Oschkinat H, Mayr I, Huber R, Angliker H, Stone SR, Bode W. The interaction of thrombin with fibrinogen. A structural basis for its specificity. Eur J Biochem 1992; 206: 187-95.
20 Mathews II, Padmanabhan KP, Ganesh V, Tulinsky A, Ishii M, Chen J, Turck CW, Coughlin SR, Fenton JW. Crystallographic structures of thrombin complexed with throm-bin receptor peptides: existence of expected and novel binding modes. Biochemistry 1994; 33: 3266-79.
21 Hrabal R, Komives EA, Ni F. Structural resiliency of an EGF-like subdomain bound to its target protein, thrombin. Protein Sci 1996; 5: 195-203.
22 Bode W, Mayr I, Baumann U, Huber R, Stone SR, Hofsteenge J. The refined 1.9 Å crystal structure of human alpha-thrombin: interaction with D-Phe-Pro-Arg chloromethylketone and significance of the Tyr-Pro-Pro-Trp insertion segment. EMBO J 1989; 8: 3467-75.
23 Kohler HP, Grant PJ. The role of factor XIIIVal34Leu in cardiovascular disease. Quart J Med 1999; 92: 67-72.
24 Ni F. Recent Developments in Transferred NOE Methods. Prog. NMR Spec 1994; 26: 517-606.
25 Campbell AP, Sykes BD. The two-dimensional transferred nuclear Overhauser effect: theory and practice. Annu. Rev. Biophys Biomol Struct 1993; 22: 99-122.