Comparison of the Actions of Thrombin and the Thrombin-Like Venom Enzymes Ancrod and Batroxobin

Dl, Aronson

doi:10.1055/s-0038-1648004

Cited by 44 publications

(31 citation statements)

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“…In contrast, clot accretion induced by batroxobin is similar regardless of whether batroxobin is bound to ␥ A /␥ A -or ␥ A /␥Ј-fibrin clots (p ϭ 0.18). It is notable that clot accretion induced by fibrinbound batroxobin is ϳ18-fold greater than that induced by fibrin-bound thrombin (p Ͻ 0.001); this indicates that the procoagulant activity of fibrin-bound batroxobin is greater than that of fibrin-bound thrombin, likely reflecting the fact that batroxobin is not inhibited by antithrombin or heparin cofactor II (20).…”

Section: Thrombin Competition Of Batroxobin Binding To Fibrin(ogen)-mentioning

confidence: 94%

“…Notably, batroxobin lacks the 60-and ␥-loops and the Na ϩ -binding site, domains that endow thrombin with its specificity for a wide repertoire of macromolecular substrates (37,41). The absence of these domains may explain why fibrinogen is the sole substrate of batroxobin (20). Without the 60-and ␥-loops and the Na ϩ -binding site, the interaction of batroxobin with fibrinogen is likely to depend on other residues in the vicinity of the active site of batroxobin (42,43).…”

Section: Discussionmentioning

confidence: 99%

“…In contrast to thrombin, fibrinogen is the sole substrate for batroxobin, and batroxobin only releases FpA and is not inhibited by antithrombin or heparin cofactor II (20). Because of these properties, batroxobin is often used in clinical laboratories to determine whether prolonged thrombin clotting times are the result of heparin contamination or abnormal fibrinogen molecules (21).…”

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confidence: 99%

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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: Thrombin Competition Of Batroxobin Binding To Fibrin(ogen)-mentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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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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“…4, C and D). These serine proteinases are thrombin-like enzymes, which make blood unclottable, and it is safe to state that type A venom contains different serine proteinases affecting either ␣ or ␤ chain because it is unlikely that the same serine proteinases could be affecting both chains (63,64). A combination of both metallo-and serine proteinases exist in a higher abundance in type B venom than in type A venom (Table II), and thus, the combination of both these proteinases, in particularly the effect of ␣ chain cleavage by metalloproteinases, could very well be the result of the delayed clotting time of blood treated with type B venom (Fig.…”

Section: Application Of the Sopil Methods To The Quantitative Analysismentioning

confidence: 99%

Quantitative Analysis of Snake Venoms Using Soluble Polymer-based Isotope Labeling

Galán

Guo

Sánchez

et al. 2008

Molecular & Cellular Proteomics

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We present the design and synthesis of a new quantitative strategy termed soluble polymer-based isotope labeling (SoPIL) and its application as a novel and inclusive method for the identification and relative quantification of individual proteins in complex snake venoms. The SoPIL reagent selectively captures and isolates cysteine-containing peptides, and the subsequent tagged peptides are released and analyzed using nanoflow liquid chromatography-tandem mass spectrometry. The SoPIL strategy was used to quantify venom proteins from two pairs of venomous snakes: Crotalus scutulatus scutulatus type A, C. scutulatus scutulatus type B, Crotalus oreganus helleri, and Bothrops colombiensis. The identification and accurate quantification of proteins in high throughput analysis are essential components of proteomics strategies for biomarker discovery and studying cellular functions and processes (1, 2). Although the technology to measure mRNA expression is more established, the measurement of differential protein expression provides a more direct, accurate, and versatile way to detect global changes in cellular dynamics in health and disease (3). Over the past several years, many quantitative techniques and strategies have been introduced and thoroughly examined. The traditional and frequently used method to investigate differential protein abundances on a large scale between samples from different sources is the staining of proteins separated by two-dimensional (2D) 1 PAGE. This method falls short in its reproducibility and its inability to detect low abundance and hydrophobic proteins (4). Recently label-free approaches primarily based on the use of LC and highly accurate mass spectrometers have been investigated (5, 6). These methods, however, rely heavily on computational software for data treatment.Stable isotopic labeling has remained the most popular method for quantitative proteomics. The introduction of stable isotopes has typically been carried out by (i) chemical derivatization of proteins or peptides, e.g. via the ICAT (7), isobaric tagging for relative and absolute quantitation (iTRAQ) (8), and isotope-coded protein labeling (9) methods; (ii) enzyme catalyzed labeling, e.g. proteolysis in heavy water incorporates 18 O into the newly generated carboxyl-terminal carboxyl groups (10); and (iii) metabolic labeling methods to incorporate isotopically labeled amino acid residues into proteins (stable isotope labeling using amino acids in cell culture (SI-LAC)) (11). ICAT is probably the best described chemical approach. Over the years, both advantages and disadvantages of ICAT have been recognized. Several variations and modifications of ICAT have been attempted to make it more practical and simpler, such as acid-cleavable ICAT reagents (12) and a solid phase based version (13). The adaptation of the solid phase capture and release process is a significant improvement, and the method obviates extra purification steps and has the potential for automation and high throughput experiments. A side-by-side comparison w...

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“…Proteases that hydrolyze fibrinogen are termed as snake venom fibrinogenolytic enzymes (SVFEs). SVFEs that hydrolyze fibrinogen-releasing fibrinopeptide A and/or B can directly induce fibrin clot formation and have pro-coagulant activity when tested on citrated plasma (Aronson, 1976). Such enzymes are called snake venom thrombin-like enzymes (SVTLEs).…”

Section: Introductionmentioning

confidence: 99%

Malabarase, a serine protease with anticoagulant activity from Trimeresurus malabaricus venom

Kumar

Manjunath

Joshi

et al. 2013

Comparative Biochemistry and Physiology Part B: Biochemistry an

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Comparison of the Actions of Thrombin and the Thrombin-Like Venom Enzymes Ancrod and Batroxobin

Cited by 44 publications

References 0 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

Quantitative Analysis of Snake Venoms Using Soluble Polymer-based Isotope Labeling

Malabarase, a serine protease with anticoagulant activity from Trimeresurus malabaricus venom

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