Studies on the basis for the properties offibrin produced from fibrinogen-containing chains

Department of Biomedical Sciences, College of Health Sciences, Marquette University, Schroeder Health Complex, 426, PO Box 1881, Milwaukee, WI 53233-1881, USA.
Blood (Impact Factor: 10.45). 11/2005; 106(8):2730-6. DOI: 10.1182/blood-2005-01-0240
Source: PubMed


Human fibrinogen 1 is homodimeric with respect to its gamma chains (gammaA-gammaA'), whereas fibrinogen 2 molecules each contain one gammaA (gammaA1-411V) and one gamma' chain, which differ by containing a unique C-terminal sequence from gamma'408 to 427L that binds thrombin and factor XIII. We investigated the structural and functional features of these fibrins and made several observations. First, thrombin-treated fibrinogen 2 produced finer, more branched clot networks than did fibrin 1. These known differences in network structure were attributable to delayed release of fibrinopeptide (FP) A from fibrinogen 2 by thrombin, which in turn was likely caused by allosteric changes at the thrombin catalytic site induced by thrombin exosite 2 binding to the gamma' chains. Second, cross-linking of fibrin gamma chains was virtually the same for both types of fibrin. Third, the acceleratory effect of fibrin on thrombin-mediated XIII activation was more prominent with fibrin 1 than with fibrin 2, and this was also attributable to allosteric changes at the catalytic site induced by thrombin binding to gamma' chains. Fourth, fibrinolysis of fibrin 2 was delayed compared with fibrin 1. Altogether, differences between the structure and function of fibrins 1 and 2 are attributable to the effects of thrombin binding to gamma' chains.

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    • "In order to clarify whether thrombin can still recognize the cleavage sites in the N-terminal region of fibrinogen to promote fibrinogenesis (Siebenlist et al., 2005), RCC-treated fibrinogen was incubated with thrombin and aliquots were examined by MALDI-TOF mass spectrometry. Cleavage of fibrinogen releases fibrinopeptide A and B, peptides of ∼1900 Da and ∼2400 Da in length. "
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    ABSTRACT: Fibrinogen is a key protein involved in coagulation and its deposition on blood vessel walls plays an important role in the pathology of atherosclerosis. Although the causes of fibrinogen (fibrin) deposition have been studied in depth, little is known about the relationship between fibrinogen deposition and reactive carbonyl compounds (RCCs), compounds which are produced and released into the blood and react with plasma protein especially under conditions of oxidative stress and inflammation. Here, we investigated the effect of glycolaldehyde on the activity and deposition of fibrinogen compared with the common RCCs acrolein, methylglyoxal, glyoxal and malondialdehyde. At the same concentration (1 mmol/L), glycolaldehyde and acrolein had a stronger suppressive effect on fibrinogen activation than the other three RCCs. Fibrinogen aggregated when it was respectively incubated with glycolaldehyde and the other RCCs, as demonstrated by SDS-PAGE, electron microscopy and intrinsic fluorescence intensity measurements. Staining with Congo Red showed that glycolaldehyde- and acrolein-fibrinogen distinctly formed amyloid-like aggregations. Furthermore, the five RCCs, particularly glycolaldehyde and acrolein, delayed human plasma coagulation. Only glycolaldehyde showed a markedly suppressive effect on fibrinogenesis, none did the other four RCCs when their physiological blood concentrations were employyed, respectively. Taken together, it is glycolaldehyde that suppresses fibrinogenesis and induces protein aggregation most effectively, suggesting a putative pathological process for fibrinogen (fibrin) deposition in the blood.
    Protein & Cell 07/2012; 3(8):627-40. DOI:10.1007/s13238-012-2057-y · 3.25 Impact Factor
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    • "It was also known that the c¢ chain lacked a binding site required for platelet adhesion and aggregation (Farrell et al, 1992; Kirschbaum et al, 1992), but the lack of an activity did not clarify the physiological role of cA/c¢ fibrinogen. More recent evidence has shown that cA/c¢ fibrinogen forms fibrin clots that are more rapidly cross-linked by factor XIIIa and are resistant to fibrinolysis (Falls & Farrell, 1997), although other investigators have shown conflicting evidence that cA/c¢ fibrin is less rapidly cross-linked (Siebenlist et al, 2005). In addition, studies have shown that the c¢ chain binds directly to the "
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    ABSTRACT: The minor gammaA/gamma' isoform of fibrinogen contains a high affinity binding site for thrombin exosite II that is lacking in the major fibrinogen isoform, gammaA/gammaA fibrinogen. The biological consequences of gamma' chain binding to thrombin were therefore investigated. Coagulation assays, thrombin activity assays, and a primate thrombosis model were used to characterize the biological effects of the gamma' 410-427 peptide. The gamma' peptide had little effect on thrombin cleavage of the small peptidyl substrate tosyl-glycyl-prolyl-arginine-4-nitranilide acetate. However, in vitro assays demonstrated that the gamma' peptide inhibited thrombin cleavage of larger proteinaceous substrates, including fibrinogen and factor VIII. The gamma' peptide inhibited the activated partial thromboplastin time in plasma and showed greater inhibition of activated partial thromboplastin time assays than prothrombin time assays, consistent with the inhibition of factor VIII cleavage. Studies in a baboon thrombosis model showed that the gamma' 410-427 peptide inhibited fibrin-rich thrombus formation (typical of venous thrombi) and, to a lesser extent, platelet-rich thrombus formation (typical of arterial thrombi). These results indicate that binding of thrombin exosite II by the gamma' peptide has selective effects on the intrinsic pathway.
    British Journal of Haematology 12/2007; 139(3):494-503. DOI:10.1111/j.1365-2141.2007.06825.x · 4.71 Impact Factor
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