Characterization of a new platelet aggregating factor from crotoxin Crotalus durissus cascavella venom

Página do item simplificado
dc.contributor.authorFonseca F.V.
dc.contributor.authorAntunes E.
dc.contributor.authorMorganti R.P.
dc.contributor.authorMonteiro H.S.A.
dc.contributor.authorMartins A.M.C.
dc.contributor.authorToyama D.O.
dc.contributor.authorMarangoni S.
dc.contributor.authorToyama M.H.
dc.date.accessioned2024-03-13T01:43:13Z
dc.date.available2024-03-13T01:43:13Z
dc.date.issued2006
dc.description.abstractIn this article we investigated the platelet aggregating activity of whole crotoxin and its subunits isolated from Crotalus durissus cascavella venom. During the purification protocols of the venom, using HPLC molecular exclusion, we detected the presence of two different serine protease activities in the gyroxin fraction, and another in the crotoxin fraction, which induced strong and irreversible platelet aggregation, in addition to blood coagulation. From crotoxin, we isolated PLA2, crotapotin (both fractions corresponding approximately 85% of whole crotoxin) and another minor fraction (F20) that exhibited serine protease activity. After a new fractionation on reverse phase HPLC chromatography, we obtained three other fractions named as F201, F202 and F203. F202 was obtained with high degree of molecular homogeneity with molecular mass of approximately 28 kDa and a high content of acidic amino residues, such as aspartic acid and glutamic acid. Other important amino acids were histidine, cysteine and lysine. This protein exhibited a high specificity for BApNA, a Michaelis-Menten behavior with Vmax estimated in 5.64 μM/min and a Km value of 0.58 mM for this substrate. In this work, we investigated the ability of F202 to degrade fibrinogen and observed α and β chain cleavage. Enzymatic as well as the platelet aggregation activities were strongly inhibited when incubated with TLCK and PMSF, specific inhibitors of serine protease. Also, F202 induced platelet aggregation in washed and platelet-rich plasma, and in both cases, TLCK inhibited its activity. The N-terminal amino acid sequence of F202 presented a high amino acid sequence homology with other thrombin-like proteins, but it was significantly different from gyroxin. These results showed that crotoxin is a highly heterogeneous protein composed of PLA2, thrombin-like and other fractions that might explain the diversity of physiological and pharmacological activities of this protein. © 2006 Springer Science+Business Media, Inc.
dc.description.firstpage183
dc.description.issuenumber3
dc.description.lastpage192
dc.description.volume25
dc.identifier.doi10.1007/s10930-006-9001-z
dc.identifier.issn1875-8355
dc.identifier.urihttps://dspace.mackenzie.br/handle/10899/37829
dc.relation.ispartofProtein Journal
dc.rightsAcesso Restrito
dc.subject.otherlanguageCrotalus durissus cascavella
dc.subject.otherlanguagePlatelet aggregation
dc.subject.otherlanguageSnake venom
dc.subject.otherlanguageThrombin like
dc.titleCharacterization of a new platelet aggregating factor from crotoxin Crotalus durissus cascavella venom
dc.typeArtigo
local.scopus.citations21
local.scopus.eid2-s2.0-33746798461
local.scopus.updated2024-05-01
local.scopus.urlhttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=33746798461&origin=inward
Arquivos
Coleções
Artigos de periódico