Crotalase, a Fibrinogen-Clotting Snake Venom Enzyme: Primary Structure and Evidence for a Fibrinogen Recognition Exosite Different from Thrombin

Henschen-Edman, Agnes H.; Theodor, Ida; Edwards, Brian F.P.; Pirkle, Hubert

doi:10.1055/s-0037-1614423

Cited by 32 publications

(18 citation statements)

References 43 publications

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“…In particular, Tyr-76 of thrombin provides the anchor for a hydrophobic patch surrounded by basic residues that interact with a complementary region on fibrinogen (39,40). Although only 50% of the corresponding residues are retained, a homologous fibrinogen recognition exosite in batroxobin has been proposed (43,44). The observation that the interaction of batroxobin with ␥ A /␥ A -fibrin clots is less readily disrupted by high salt than that of thrombin raises the possibility that hydrophobic interactions may be more important for batroxobin binding to the fibrinogen recognition exosite than they are for thrombin binding.…”

Section: Discussionmentioning

confidence: 99%

Batroxobin Binds Fibrin with Higher Affinity and Promotes Clot Expansion to a Greater Extent than Thrombin

Stafford

Leslie

et al. 2013

Journal of Biological Chemistry

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Batroxobin is a thrombin-like serine protease from the venom of Bothrops atrox moojeni that clots fibrinogen. In contrast to thrombin, which releases fibrinopeptide A and B from the NH2-terminal domains of the Aα- and Bβ-chains of fibrinogen, respectively, batroxobin only releases fibrinopeptide A. Because the mechanism responsible for these differences is unknown, we compared the interactions of batroxobin and thrombin with the predominant γA/γA isoform of fibrin(ogen) and the γA/γ′ variant with an extended γ-chain. Thrombin binds to the γ′-chain and forms a higher affinity interaction with γA/γ′-fibrin(ogen) than γA/γA-fibrin(ogen). In contrast, batroxobin binds both fibrin(ogen) isoforms with similar high affinity (Kd values of about 0.5 μm) even though it does not interact with the γ′-chain. The batroxobin-binding sites on fibrin(ogen) only partially overlap with those of thrombin because thrombin attenuates, but does not abrogate, the interaction of γA/γA-fibrinogen with batroxobin. Furthermore, although both thrombin and batroxobin bind to the central E-region of fibrinogen with a Kd value of 2–5 μm, the α(17–51) and Bβ(1–42) regions bind thrombin but not batroxobin. Once bound to fibrin, the capacity of batroxobin to promote fibrin accretion is 18-fold greater than that of thrombin, a finding that may explain the microvascular thrombosis that complicates envenomation by B. atrox moojeni. Therefore, batroxobin binds fibrin(ogen) in a manner distinct from thrombin, which may contribute to its higher affinity interaction, selective fibrinopeptide A release, and prothrombotic properties.

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Section: Discussionmentioning

confidence: 99%

Batroxobin Binds Fibrin with Higher Affinity and Promotes Clot Expansion to a Greater Extent than Thrombin

Stafford

Leslie

et al. 2013

Journal of Biological Chemistry

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“…According to LM-TL and crotalase models, the residues that can comprise this region are Phe95, Trp99, Tyr172, Phe174, Leu176, Phe214 and Trp215, which are spatially analogous to Tyr60A, Trp60D, segment 97 -99, Ile174 and Trp215 of thrombin ( fig. 2 B) [52,53,73]. This region seems to be represented in some SVTLEs, though not all positions are conserved in this subfamily such as Phe95, 174 and 214 [74] ( fig.…”

Section: Catalytic and Binding Sitesmentioning

confidence: 97%

“…Numbering is according to chymotrypsinogen [79]. for stabilization of the snake venom serine protease structure in studies of TSV-PA [51], and three-dimensional (3D) models of LM-TL from Lachesis muta [52], crotalase from Crotalus adamanteus [53] and other SVTLEs [54,55]. The secondary structure prediction for SVTLE sequence reveals several b-sheet structures, inferring a hypothetical b/b hydrolase fold typical of serine proteases from the chymotrypsin family (figs 1, 2 A) [56,57].…”

Section: Common Features Of Svtlesmentioning

confidence: 99%

Snake venom thrombin-like enzymes: from reptilase to now

Castro

Zingali

Albuquerque

et al. 2004

Cellular and Molecular Life Sciences (CMLS)

170

123

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The snake venom thrombin-like enzymes (SVTLEs) comprise a number of serine proteases functionally and structurally related to thrombin. Until recently, only nine complete sequences of this subgroup of the serine protease family were known. Over the past 5 years, the primary structure of several SVTLEs has been characterized, and now this family includes several members. Of particular interest is their possible use in pathologies such as thrombosis. The aim of the present review is to summarize the state of the art concerning the evolutionary, structural and biological features of the SVTLEs.

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“…On the basis of a model, an alternative Fg recognition site was proposed for crotalase [20,34]. Given that a number of basic and hydrophobic residues are present in SVTLEs at a region equivalent to the ESI in thrombin, the alternative site seems to be rather an extension of the ESI specific to SVTLEs, that includes Lys84a and Lys86.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

Molecular Basis for the Partition of the Essential Functions of Thrombin among Snake Venom Serine Proteinases

Maroun

2001

Pathophysiol Haemos Thromb

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Thrombin is a mammalian serine proteinase that plays a prominent role in the maintenance and regulation of hemostasis through its interaction with various substrates and/or ligands. The venoms of several snakes contain glycosylated serine proteinases that have been recognized to possess one or more of the essential activities of thrombin on fibrinogen (Fg) and/or platelets. These proteinases share about 60% sequence identity. One class of snake venom serine proteinases are those known as thrombin-like (TLE), named after their ability to directly clot Fg in order to preferentially produce fibrinopeptide A, fibrinopeptide B or both. To understand the molecular basis of this phenomenon, the corresponding amino acid sequences and molecular structures need to be analyzed. Given the absence of experimentally determined tertiary structures of snake venom, TLEs, three-dimensional molecular models should prove useful in this context. Towards this goal, we obtained models of snake venom TLEs that used TSV-PA as template, TSV-PA being the only snake venom serine proteinase whose crystal structure is known to date. Along with a comparative sequence analysis the models contribute to the identification and description of thrombin-homologous or alternative binding sites, helping thus to understand differences in specificity.

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Crotalase, a Fibrinogen-Clotting Snake Venom Enzyme: Primary Structure and Evidence for a Fibrinogen Recognition Exosite Different from Thrombin

Cited by 32 publications

References 43 publications

Batroxobin Binds Fibrin with Higher Affinity and Promotes Clot Expansion to a Greater Extent than Thrombin

Batroxobin Binds Fibrin with Higher Affinity and Promotes Clot Expansion to a Greater Extent than Thrombin

Snake venom thrombin-like enzymes: from reptilase to now

Molecular Basis for the Partition of the Essential Functions of Thrombin among Snake Venom Serine Proteinases

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