Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography

Detalhes bibliográficos
Autor(a) principal: Canduri, F.
Data de Publicação: 1998
Outros Autores: Ward, R. J., de Azevedo, W. F., Gomes, RAS, Arni, R. K.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1080/15216549800203222
http://hdl.handle.net/11449/38170
Resumo: Cathepsin D, a lysosomal aspartic protease, has been purified from porcine liver using a combination of pepstatin-A agarose and Affi-Gel Blue affinity chromatography, followed by size-exclusion chromatography. The purified protein consists of two polypeptide chains of 15 and 30 kDa, and has an isoelectric point of 6.8. Porcine liver cathepsin D has maximum activity at pH 2.5-3.0 as determined by its activity against hemoglobin, with a K-cat of 14.3 s(-1) and a k(cat)/K-M of 2.70 x 10(6) s(-1) M-1 as determined by the hydrolysis of a fluorogenic peptide substrate.
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spelling Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatographycathepsin Daspartic proteaselysosomal enzymeaffinity chromatographypepstatin ACathepsin D, a lysosomal aspartic protease, has been purified from porcine liver using a combination of pepstatin-A agarose and Affi-Gel Blue affinity chromatography, followed by size-exclusion chromatography. The purified protein consists of two polypeptide chains of 15 and 30 kDa, and has an isoelectric point of 6.8. Porcine liver cathepsin D has maximum activity at pH 2.5-3.0 as determined by its activity against hemoglobin, with a K-cat of 14.3 s(-1) and a k(cat)/K-M of 2.70 x 10(6) s(-1) M-1 as determined by the hydrolysis of a fluorogenic peptide substrate.UNESP, IBILCE, Dept Fis, BR-15054000 Sao Jose do Rio Preto, SP, BrazilFac Med Triangulo Mineiro, Dept Bioquim Celular & Biofis, BR-38015050 Uberaba, MG, BrazilUNESP, IBILCE, Dept Fis, BR-15054000 Sao Jose do Rio Preto, SP, BrazilAcademic Press AustUniversidade Estadual Paulista (Unesp)Fac Med Triangulo MineiroCanduri, F.Ward, R. J.de Azevedo, W. F.Gomes, RASArni, R. K.2014-05-20T15:28:21Z2014-05-20T15:28:21Z1998-07-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article797-803application/pdfhttp://dx.doi.org/10.1080/15216549800203222Biochemistry and Molecular Biology International. Marrickville: Academic Press Aust, v. 45, n. 4, p. 797-803, 1998.1039-9712http://hdl.handle.net/11449/3817010.1080/15216549800203222WOS:000075300600019WOS000075300600019.pdf91625089789458870000-0003-2460-1145Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBiochemistry and Molecular Biology Internationalinfo:eu-repo/semantics/openAccess2023-11-20T06:13:44Zoai:repositorio.unesp.br:11449/38170Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T18:15:13.553973Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography
title Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography
spellingShingle Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography
Canduri, F.
cathepsin D
aspartic protease
lysosomal enzyme
affinity chromatography
pepstatin A
title_short Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography
title_full Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography
title_fullStr Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography
title_full_unstemmed Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography
title_sort Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography
author Canduri, F.
author_facet Canduri, F.
Ward, R. J.
de Azevedo, W. F.
Gomes, RAS
Arni, R. K.
author_role author
author2 Ward, R. J.
de Azevedo, W. F.
Gomes, RAS
Arni, R. K.
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
Fac Med Triangulo Mineiro
dc.contributor.author.fl_str_mv Canduri, F.
Ward, R. J.
de Azevedo, W. F.
Gomes, RAS
Arni, R. K.
dc.subject.por.fl_str_mv cathepsin D
aspartic protease
lysosomal enzyme
affinity chromatography
pepstatin A
topic cathepsin D
aspartic protease
lysosomal enzyme
affinity chromatography
pepstatin A
description Cathepsin D, a lysosomal aspartic protease, has been purified from porcine liver using a combination of pepstatin-A agarose and Affi-Gel Blue affinity chromatography, followed by size-exclusion chromatography. The purified protein consists of two polypeptide chains of 15 and 30 kDa, and has an isoelectric point of 6.8. Porcine liver cathepsin D has maximum activity at pH 2.5-3.0 as determined by its activity against hemoglobin, with a K-cat of 14.3 s(-1) and a k(cat)/K-M of 2.70 x 10(6) s(-1) M-1 as determined by the hydrolysis of a fluorogenic peptide substrate.
publishDate 1998
dc.date.none.fl_str_mv 1998-07-01
2014-05-20T15:28:21Z
2014-05-20T15:28:21Z
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/10.1080/15216549800203222
Biochemistry and Molecular Biology International. Marrickville: Academic Press Aust, v. 45, n. 4, p. 797-803, 1998.
1039-9712
http://hdl.handle.net/11449/38170
10.1080/15216549800203222
WOS:000075300600019
WOS000075300600019.pdf
9162508978945887
0000-0003-2460-1145
url http://dx.doi.org/10.1080/15216549800203222
http://hdl.handle.net/11449/38170
identifier_str_mv Biochemistry and Molecular Biology International. Marrickville: Academic Press Aust, v. 45, n. 4, p. 797-803, 1998.
1039-9712
10.1080/15216549800203222
WOS:000075300600019
WOS000075300600019.pdf
9162508978945887
0000-0003-2460-1145
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Biochemistry and Molecular Biology International
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 797-803
application/pdf
dc.publisher.none.fl_str_mv Academic Press Aust
publisher.none.fl_str_mv Academic Press Aust
dc.source.none.fl_str_mv Web of Science
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_ 1808128911614148608

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