Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions

Detalhes bibliográficos
Autor(a) principal: de Oliveira, Laudicéia Alves [UNESP]
Data de Publicação: 2017
Outros Autores: Ferreira, Rui Seabra [UNESP], Barraviera, Benedito [UNESP], de Carvalho, Francilene Capel Tavares [UNESP], de Barros, Luciana Curtolo [UNESP], dos Santos, Lucilene Delazari [UNESP], Pimenta, Daniel Carvalho [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1186/s40409-017-0136-5
http://hdl.handle.net/11449/175556
Resumo: Background: Classically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported. Methods: The crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing. Results: The RP-HPLC profile of the isolated crotapotin chains already indicated that the α chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses. Conclusion: It was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the α chain, at positions 31 and 40. Moreover, substitutions could also be observed in β and γ chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.
id UNSP_2fd96d5b9bdbd8db0959990c3de8a78f
oai_identifier_str oai:repositorio.unesp.br:11449/175556
network_acronym_str UNSP
network_name_str Repositório Institucional da UNESP
repository_id_str 2946
spelling Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutionsCrotalus durissus terrificusCrotapotinCrotoxinIsoformsVenomBackground: Classically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported. Methods: The crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing. Results: The RP-HPLC profile of the isolated crotapotin chains already indicated that the α chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses. Conclusion: It was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the α chain, at positions 31 and 40. Moreover, substitutions could also be observed in β and γ chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.Sao Paulo State University (UNESP) Postgraduate Program in Tropical Diseases Botucatu Medical SchoolSão Paulo State University (UNESP) Center for the Studies of Venoms and Venomous Animals (CEVAP)Butantan Institute Laboratory of Biochemistry and Biophysics, Av. Vital Brazil, 1500Sao Paulo State University (UNESP) Postgraduate Program in Tropical Diseases Botucatu Medical SchoolSão Paulo State University (UNESP) Center for the Studies of Venoms and Venomous Animals (CEVAP)Universidade Estadual Paulista (Unesp)Laboratory of Biochemistry and Biophysicsde Oliveira, Laudicéia Alves [UNESP]Ferreira, Rui Seabra [UNESP]Barraviera, Benedito [UNESP]de Carvalho, Francilene Capel Tavares [UNESP]de Barros, Luciana Curtolo [UNESP]dos Santos, Lucilene Delazari [UNESP]Pimenta, Daniel Carvalho [UNESP]2018-12-11T17:16:17Z2018-12-11T17:16:17Z2017-11-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1186/s40409-017-0136-5Journal of Venomous Animals and Toxins Including Tropical Diseases, v. 23, n. 1, 2017.1678-91991678-9180http://hdl.handle.net/11449/17555610.1186/s40409-017-0136-5S1678-919920170001003192-s2.0-85035104772S1678-91992017000100319.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Venomous Animals and Toxins Including Tropical Diseases0,573info:eu-repo/semantics/openAccess2024-08-15T15:23:28Zoai:repositorio.unesp.br:11449/175556Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-15T15:23:28Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions
title Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions
spellingShingle Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions
de Oliveira, Laudicéia Alves [UNESP]
Crotalus durissus terrificus
Crotapotin
Crotoxin
Isoforms
Venom
title_short Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions
title_full Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions
title_fullStr Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions
title_full_unstemmed Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions
title_sort Crotalus durissus terrificus crotapotin naturally displays preferred positions for amino acid substitutions
author de Oliveira, Laudicéia Alves [UNESP]
author_facet de Oliveira, Laudicéia Alves [UNESP]
Ferreira, Rui Seabra [UNESP]
Barraviera, Benedito [UNESP]
de Carvalho, Francilene Capel Tavares [UNESP]
de Barros, Luciana Curtolo [UNESP]
dos Santos, Lucilene Delazari [UNESP]
Pimenta, Daniel Carvalho [UNESP]
author_role author
author2 Ferreira, Rui Seabra [UNESP]
Barraviera, Benedito [UNESP]
de Carvalho, Francilene Capel Tavares [UNESP]
de Barros, Luciana Curtolo [UNESP]
dos Santos, Lucilene Delazari [UNESP]
Pimenta, Daniel Carvalho [UNESP]
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
Laboratory of Biochemistry and Biophysics
dc.contributor.author.fl_str_mv de Oliveira, Laudicéia Alves [UNESP]
Ferreira, Rui Seabra [UNESP]
Barraviera, Benedito [UNESP]
de Carvalho, Francilene Capel Tavares [UNESP]
de Barros, Luciana Curtolo [UNESP]
dos Santos, Lucilene Delazari [UNESP]
Pimenta, Daniel Carvalho [UNESP]
dc.subject.por.fl_str_mv Crotalus durissus terrificus
Crotapotin
Crotoxin
Isoforms
Venom
topic Crotalus durissus terrificus
Crotapotin
Crotoxin
Isoforms
Venom
description Background: Classically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported. Methods: The crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing. Results: The RP-HPLC profile of the isolated crotapotin chains already indicated that the α chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses. Conclusion: It was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the α chain, at positions 31 and 40. Moreover, substitutions could also be observed in β and γ chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.
publishDate 2017
dc.date.none.fl_str_mv 2017-11-28
2018-12-11T17:16:17Z
2018-12-11T17:16:17Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1186/s40409-017-0136-5
Journal of Venomous Animals and Toxins Including Tropical Diseases, v. 23, n. 1, 2017.
1678-9199
1678-9180
http://hdl.handle.net/11449/175556
10.1186/s40409-017-0136-5
S1678-91992017000100319
2-s2.0-85035104772
S1678-91992017000100319.pdf
url http://dx.doi.org/10.1186/s40409-017-0136-5
http://hdl.handle.net/11449/175556
identifier_str_mv Journal of Venomous Animals and Toxins Including Tropical Diseases, v. 23, n. 1, 2017.
1678-9199
1678-9180
10.1186/s40409-017-0136-5
S1678-91992017000100319
2-s2.0-85035104772
S1678-91992017000100319.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Journal of Venomous Animals and Toxins Including Tropical Diseases
0,573
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
_version_ 1808128211532382208