Expression and purification of untagged Glnk proteins from actinobacteria

Autores
Gerhardt, Edileusa C. M.; Moure, Vivian R.; Souza, Andrey W.; Pedrosa, Fabio O.; Souza, Emanuel M.; Diacovich, Lautaro; Gramajo, Hugo Cesar; Huergo, Luciano F.
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The PII protein family constitutes one of the most conserved and well distributed family of signal transduction proteins in nature. These proteins play key roles in nitrogen and carbon metabolism. PII function has been welldocumented in Gram-negative bacteria. However, there are very few reports describing the in vitro properties and function of PII derived from Gram-positive bacteria. Here we present the heterologous expression and efficientpurification protocols for untagged PII from three Actinobacteria of medical and biotechnological interest namely: Mycobacterium tuberculosis, Rhodococcus jostii and Streptomyces coelicolor. Circular dichroism and gel filtrationanalysis supported that the purified proteins are correctly folded. The purification protocol described here will facilitate biochemical studies and help to uncover the biochemical functions of PII proteins in Actinobacteria.
Fil: Gerhardt, Edileusa C. M.. Universidade Federal do Paraná; Brasil
Fil: Moure, Vivian R.. Universidade Federal do Paraná; Brasil
Fil: Souza, Andrey W.. Universidade Federal do Paraná; Brasil
Fil: Pedrosa, Fabio O.. Universidade Federal do Paraná; Brasil
Fil: Souza, Emanuel M.. Universidade Federal do Paraná; Brasil
Fil: Diacovich, Lautaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Gramajo, Hugo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Huergo, Luciano F.. Universidade Federal do Paraná; Brasil
Materia
PII PROTEINS
ACTINOBACTERIA
NITROGEN
METABOLISM
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/53368
Acceder
id CONICETDig_25a937c49bdb8e75825087b56af3fdc8
oai_identifier_str oai:ri.conicet.gov.ar:11336/53368
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Expression and purification of untagged Glnk proteins from actinobacteriaGerhardt, Edileusa C. M.Moure, Vivian R.Souza, Andrey W.Pedrosa, Fabio O.Souza, Emanuel M.Diacovich, LautaroGramajo, Hugo CesarHuergo, Luciano F.PII PROTEINSACTINOBACTERIANITROGENMETABOLISMhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The PII protein family constitutes one of the most conserved and well distributed family of signal transduction proteins in nature. These proteins play key roles in nitrogen and carbon metabolism. PII function has been welldocumented in Gram-negative bacteria. However, there are very few reports describing the in vitro properties and function of PII derived from Gram-positive bacteria. Here we present the heterologous expression and efficientpurification protocols for untagged PII from three Actinobacteria of medical and biotechnological interest namely: Mycobacterium tuberculosis, Rhodococcus jostii and Streptomyces coelicolor. Circular dichroism and gel filtrationanalysis supported that the purified proteins are correctly folded. The purification protocol described here will facilitate biochemical studies and help to uncover the biochemical functions of PII proteins in Actinobacteria.Fil: Gerhardt, Edileusa C. M.. Universidade Federal do Paraná; BrasilFil: Moure, Vivian R.. Universidade Federal do Paraná; BrasilFil: Souza, Andrey W.. Universidade Federal do Paraná; BrasilFil: Pedrosa, Fabio O.. Universidade Federal do Paraná; BrasilFil: Souza, Emanuel M.. Universidade Federal do Paraná; BrasilFil: Diacovich, Lautaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Gramajo, Hugo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Huergo, Luciano F.. Universidade Federal do Paraná; BrasilLeibniz Research Centre for Working Environment and Human Factors2017-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/53368Gerhardt, Edileusa C. M.; Moure, Vivian R.; Souza, Andrey W.; Pedrosa, Fabio O.; Souza, Emanuel M.; et al.; Expression and purification of untagged Glnk proteins from actinobacteria; Leibniz Research Centre for Working Environment and Human Factors; Excli Journal; 16; 6-2017; 949-9581611-2156CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5579400/info:eu-repo/semantics/altIdentifier/url/http://www.excli.de/volume16.phpinfo:eu-repo/semantics/altIdentifier/url/http://www.excli.de/vol16/Huergo_27062017_proof.pdfinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-02-26T10:02:11Zoai:ri.conicet.gov.ar:11336/53368instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982026-02-26 10:02:11.625CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Expression and purification of untagged Glnk proteins from actinobacteria
title Expression and purification of untagged Glnk proteins from actinobacteria
spellingShingle Expression and purification of untagged Glnk proteins from actinobacteria
Gerhardt, Edileusa C. M.
PII PROTEINS
ACTINOBACTERIA
NITROGEN
METABOLISM
title_short Expression and purification of untagged Glnk proteins from actinobacteria
title_full Expression and purification of untagged Glnk proteins from actinobacteria
title_fullStr Expression and purification of untagged Glnk proteins from actinobacteria
title_full_unstemmed Expression and purification of untagged Glnk proteins from actinobacteria
title_sort Expression and purification of untagged Glnk proteins from actinobacteria
dc.creator.none.fl_str_mv Gerhardt, Edileusa C. M.
Moure, Vivian R.
Souza, Andrey W.
Pedrosa, Fabio O.
Souza, Emanuel M.
Diacovich, Lautaro
Gramajo, Hugo Cesar
Huergo, Luciano F.
author Gerhardt, Edileusa C. M.
author_facet Gerhardt, Edileusa C. M.
Moure, Vivian R.
Souza, Andrey W.
Pedrosa, Fabio O.
Souza, Emanuel M.
Diacovich, Lautaro
Gramajo, Hugo Cesar
Huergo, Luciano F.
author_role author
author2 Moure, Vivian R.
Souza, Andrey W.
Pedrosa, Fabio O.
Souza, Emanuel M.
Diacovich, Lautaro
Gramajo, Hugo Cesar
Huergo, Luciano F.
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv PII PROTEINS
ACTINOBACTERIA
NITROGEN
METABOLISM
topic PII PROTEINS
ACTINOBACTERIA
NITROGEN
METABOLISM
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The PII protein family constitutes one of the most conserved and well distributed family of signal transduction proteins in nature. These proteins play key roles in nitrogen and carbon metabolism. PII function has been welldocumented in Gram-negative bacteria. However, there are very few reports describing the in vitro properties and function of PII derived from Gram-positive bacteria. Here we present the heterologous expression and efficientpurification protocols for untagged PII from three Actinobacteria of medical and biotechnological interest namely: Mycobacterium tuberculosis, Rhodococcus jostii and Streptomyces coelicolor. Circular dichroism and gel filtrationanalysis supported that the purified proteins are correctly folded. The purification protocol described here will facilitate biochemical studies and help to uncover the biochemical functions of PII proteins in Actinobacteria.
Fil: Gerhardt, Edileusa C. M.. Universidade Federal do Paraná; Brasil
Fil: Moure, Vivian R.. Universidade Federal do Paraná; Brasil
Fil: Souza, Andrey W.. Universidade Federal do Paraná; Brasil
Fil: Pedrosa, Fabio O.. Universidade Federal do Paraná; Brasil
Fil: Souza, Emanuel M.. Universidade Federal do Paraná; Brasil
Fil: Diacovich, Lautaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Gramajo, Hugo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Huergo, Luciano F.. Universidade Federal do Paraná; Brasil
description The PII protein family constitutes one of the most conserved and well distributed family of signal transduction proteins in nature. These proteins play key roles in nitrogen and carbon metabolism. PII function has been welldocumented in Gram-negative bacteria. However, there are very few reports describing the in vitro properties and function of PII derived from Gram-positive bacteria. Here we present the heterologous expression and efficientpurification protocols for untagged PII from three Actinobacteria of medical and biotechnological interest namely: Mycobacterium tuberculosis, Rhodococcus jostii and Streptomyces coelicolor. Circular dichroism and gel filtrationanalysis supported that the purified proteins are correctly folded. The purification protocol described here will facilitate biochemical studies and help to uncover the biochemical functions of PII proteins in Actinobacteria.
publishDate 2017
dc.date.none.fl_str_mv 2017-06
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/53368
Gerhardt, Edileusa C. M.; Moure, Vivian R.; Souza, Andrey W.; Pedrosa, Fabio O.; Souza, Emanuel M.; et al.; Expression and purification of untagged Glnk proteins from actinobacteria; Leibniz Research Centre for Working Environment and Human Factors; Excli Journal; 16; 6-2017; 949-958
1611-2156
CONICET Digital
CONICET
url http://hdl.handle.net/11336/53368
identifier_str_mv Gerhardt, Edileusa C. M.; Moure, Vivian R.; Souza, Andrey W.; Pedrosa, Fabio O.; Souza, Emanuel M.; et al.; Expression and purification of untagged Glnk proteins from actinobacteria; Leibniz Research Centre for Working Environment and Human Factors; Excli Journal; 16; 6-2017; 949-958
1611-2156
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5579400/
info:eu-repo/semantics/altIdentifier/url/http://www.excli.de/volume16.php
info:eu-repo/semantics/altIdentifier/url/http://www.excli.de/vol16/Huergo_27062017_proof.pdf
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Leibniz Research Centre for Working Environment and Human Factors
publisher.none.fl_str_mv Leibniz Research Centre for Working Environment and Human Factors
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1858304967043973120
score 13.176822

Publicaciones similares