The sequence gamma-(312-324) is a fibrin-specific epitope

Schielen, WJ; Adams, HP; Leuven, K van; Voskuilen, M; Tesser, GI; Nieuwenhuizen, W

doi:10.1182/blood.v77.10.2169.bloodjournal77102169

Cited by 31 publications

(44 citation statements)

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“…FCB-2 and FCB-5 are soluble substitutes for solid-phase fibrin and can accelerate t-PA mediated activation of plasminogen. Monoclonal antibodies against a part of FCB-2 (Act-( 148-160)] [32] or FCB-5 (y-(312-325)] [33] react with fibrin, but not with fibrinogen. Moreover, Aa-Lys157 of FCB-2 and y-Lys321 of FCB-5 bind to the lysine binding sites (LBS) of plasminogen and t-PA [27,28].…”

Section: Discussionmentioning

confidence: 99%

Effects of fibrin and α2-antiplasmin on plasminogen activation by staphylokinase

et al. 1996

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Staphylokinase obtains plasminogen activating activity by forming a complex with plas-minogen. Although the enzymatic activity of staphylokinase is enhanced by fibrin, how fibrin enhances enzymatic activity has not been determined yet. The effects of fibrin, or fibrinogen fragments, on the activation of plasminogen by staphylokinase was investigated using CNBr-digested fibrinogen fragments (FCB-2 and FCB-5) and plasmin-degraded cross-linked fibrin fragments ((DD)E complex, DD fragments and E fragments). Kinetic analysis of the activity of staphylokinase revealed that its plasminogen activating activity, which was expressed as kcatlKm, was enhanced by FCB-2 (10-fold) and FCB-5 (5-fold). These fibrin fragments caused 38-, 30-, and 8.5-fold increases in activity for the DD fragment, (DD)E complex and E fragment, respectively. Although a,-antiplasmin inhibited the activation of plasminogen by staphylokinase, FCB-2 abolished its inhibitory effects, and the plasminogen activating activity of staphylokinase was restored. The inhibitory effects of a,-antiplasmin on the activation of mini-plasminogen by staphyloki-nase were less than for Glu-or Lys-plasminogen, and the inhibitory effect of a,-antiplasmin was not altered by fibrin or EACA. These findings indicate that the staphylokinaselplasmin-(ogen) complex reacts with fibrin even in the presence of a,-antiplasmin, and efficient plasminogen activation takes place on the surface of fibrin.

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Section: Discussionmentioning

confidence: 99%

Effects of fibrin and α2-antiplasmin on plasminogen activation by staphylokinase

et al. 1996

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“…Using monoclonal antibodies against either fibrinogen or fibrin, earlier studies have revealed that the D domains of fibrinogen and fibrin are immunologically distinct. [73][74][75] The central region of the gamma chain is inaccessible to antibodies in native fibrinogen 73,74 but hidden epitopes within it are exposed following platelet binding and/or enzyme degradation. 75,76 Finally, and directly pertinent to the present results, we find that the critically important P1 and P2 epitopes are displayed on both fibrin and surface-bound fibrinogen but are occult in native fibrinogen.…”

Section: Discussionmentioning

confidence: 99%

Molecular basis of biomaterial-mediated foreign body reactions

Eaton

Ugarova

et al. 2001

Blood

388

300

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Despite being inert and nontoxic, implanted biomaterials often trigger adverse foreign body reactions such as inflammation, fibrosis, infection, and thrombosis. With regard to the inflammatory responses to biomaterial implants, it was previously found that a crucial precedent event was the spontaneous adsorption and denaturation of fibrinogen on implant surfaces. It was further found that interactions between the phagocyte integrin Mac-1 (CD11b/CD18) and one short sequence within the fibrinogen D domain (gamma 190-202; P1) at least partially explained phagocyte accumulation on implant surfaces. However, the reason that adsorbed fibrinogen is proinflammatory--while soluble fibrinogen clearly is not--remained obscure. In this study, therefore, the question of how fibrinogen is converted to a proinflammatory state when adsorbed to biomaterial surfaces is investigated. In soluble fibrinogen, the 13 amino acid P1 sequence was found to be hidden. However, the adsorption and denaturation of fibrinogen on the surfaces of commonly used biomaterials lead to the exposure of P1 and a second neo-epitope, gamma 377-395 (P2), which also interacts with Mac-1 and is similarly occult in the soluble protein. The extent of biomaterial-mediated P1 and P2 exposure appears directly related to the severity of inflammatory responses to a test panel of biomaterials. Finally, thrombin-mediated conversion of fibrinogen to fibrin also exposes both P1 and P2 epitopes. These observations may help explain both the inflammation caused by many types of implanted biomaterials and that which occurs naturally following thrombotic events. (Blood. 2001;98:1231-1238)

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“…3,4 Fibrin degradation products, the D-D:E 1 complex and its components, dimeric D-D fragment and fragment E 1 , were prepared from plasmin digest of factor XIIIacrosslinked human fibrin. 5 Fibrin specific murine monoclonal antibodies (Mabs) against A α 148-160 and γ 312-324 regions of fibrin(ogen) 6,7 were obtained from Dr. Nieuwenhuizen. Binding studies were performed by enzyme linked immunoassay (ELISA) and by surface plasmon resonance method (SPR) using the IAsys biosensor (Fisons, Cambridge, UK) as described in References 5,8,and 9. Differential scanning calorimetry (DSC) measurements were made with DASM-4M calorimeter at a scan rate of 1 ° C/min as described elsewhere. 10 Protein concentrations varied from 1.0 to 2.0 mg/ml.…”

Section: Methodsmentioning

confidence: 99%

Conformational Changes upon Conversion of Fibrinogen into Fibrin

Medved

Tsurupa

Yakovlev

2001

Annals of the New York Academy of Sciences

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Conformational changes upon conversion of fibrinogen to fibrin result in the exposure of multiple binding sites that provide its interaction with various proteins and cells and, thus, its participation in a number of physiological and pathological processes. Here we focus on conformational changes in the fibrinogen D regions (domains) and αC‐domains that are directly involved in intermolecular interactions upon fibrin assembly. According to the current view, two αC‐domains that interact intramolecularly in fibrinogen undergo an intra‐ to intermolecular switch to form αC‐polymers in fibrin. The availability of recombinant fragments that correspond to the αC‐domain made it possible to further clarify this mechanism and to reveal novel cryptic sites in this domain for plasminogen and its activator tPA, whose exposure may play an important role in the regulation of fibrinolysis. To elucidate the mechanism of exposure of cryptic sites in the D regions, we tested the accessibility of their fibrin specific epitopes (Aα148–160 and γ312–324) that are also involved in binding of plasminogen and tPA, in several fragments derived from fibrinogen (fragment D), and crosslinked fibrin (fragment D‐D and its non‐covalent complex with the E1 fragment, D‐D:E1). Neither D nor D‐D bound tPA, plasminogen, or anti‐Aα148–160 and anti‐γ312–324 monoclonal antibodies. At the same time both epitopes became accessible in the D‐D:E1 complex. Melting of D and D‐D revealed that their domains have the same stability while in the D‐D:E1 complex they became more stable. These results indicate that upon fibrin assembly, driven primarily by the interaction between complementary binding sites of the E and two D regions, the latter undergo conformational changes that cause the exposure of their cryptic sites. They also suggest that the fibrin specific conformation of the D regions is preserved in the D‐D:E1 complex.

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The sequence gamma-(312-324) is a fibrin-specific epitope

Cited by 31 publications

References 0 publications

Effects of fibrin and α2-antiplasmin on plasminogen activation by staphylokinase

Effects of fibrin and α2-antiplasmin on plasminogen activation by staphylokinase

Molecular basis of biomaterial-mediated foreign body reactions

Conformational Changes upon Conversion of Fibrinogen into Fibrin

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