PURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASE
Autor(a) principal: | |
---|---|
Data de Publicação: | 1995 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/11600/44203 http://doi.org/10.1074/jbc.270.40.23845 |
Resumo: | Dynorphin B (Dyn B-13, also known as rimorphin) is generated from Dyn B-29 (leumorphin) by the cleavage at a single Arg residue. An enzymatic activity capable of processing at this monobasic site has been previously reported in neurosecretory vesicles of the bovine pituitary and pituitary-derived cell lines. This enzyme termed ''the dynorphin-converting enzyme'' (DCE) has been purified to apparent homogeneity from the neurointermediate lobe of the bovine pituitary using hydrophobic chromatography on phenyl-Sepharose, preparative isoelectrofocusing in a granulated gel between pH 4 to 6.5, and non-denaturing electrophoresis on 5% polyacrylamide gel, DCE exhibits a pI of about 5.1 and a molecular mass of about 54 kDa under reducing conditions. DCE is a metallopeptidase and exhibits a neutral pH optimum. Specific Inhibitors of soluble metallopeptidases such as enkephalinase (EC 3.4.24.11) or enkephalin generating neutral endopeptidase (EC 3.4.24.15) do not inhibit DCE activity indicating that DCE is distinct from these two enzymes. Cleavage site determination with matrix-assisted laser desorption ionization time of flight (MALDITOF) mass spectrometry shows that DCE cleaves the Dyn B-29 N terminus to the Arg(14) generating Dyn B-13 and Dyn B-(14-29). Among other peptides derived from Dyn B-29, DCE cleaves only those peptides that fit the predicted ''consensus motif'' for monobasic processing. These data are consistent with a broader role for the dynorphin converting enzyme in the biosynthesis of many peptide hormones and neuropeptides by processing at monobasic sites. |
id |
UFSP_2aa59419534fd35526b58cf3ab255dc9 |
---|---|
oai_identifier_str |
oai:repositorio.unifesp.br:11600/44203 |
network_acronym_str |
UFSP |
network_name_str |
Repositório Institucional da UNIFESP |
repository_id_str |
3465 |
spelling |
Berman, Y. L.Juliano, Luiz [UNIFESP]Devi, L. A.NYUUniversidade Federal de São Paulo (UNIFESP)2018-06-15T17:53:04Z2018-06-15T17:53:04Z1995-10-06Journal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 270, n. 40, p. 23845-23850, 1995.0021-9258http://repositorio.unifesp.br/11600/44203http://doi.org/10.1074/jbc.270.40.2384510.1074/jbc.270.40.23845WOS:A1995RY90900091Dynorphin B (Dyn B-13, also known as rimorphin) is generated from Dyn B-29 (leumorphin) by the cleavage at a single Arg residue. An enzymatic activity capable of processing at this monobasic site has been previously reported in neurosecretory vesicles of the bovine pituitary and pituitary-derived cell lines. This enzyme termed ''the dynorphin-converting enzyme'' (DCE) has been purified to apparent homogeneity from the neurointermediate lobe of the bovine pituitary using hydrophobic chromatography on phenyl-Sepharose, preparative isoelectrofocusing in a granulated gel between pH 4 to 6.5, and non-denaturing electrophoresis on 5% polyacrylamide gel, DCE exhibits a pI of about 5.1 and a molecular mass of about 54 kDa under reducing conditions. DCE is a metallopeptidase and exhibits a neutral pH optimum. Specific Inhibitors of soluble metallopeptidases such as enkephalinase (EC 3.4.24.11) or enkephalin generating neutral endopeptidase (EC 3.4.24.15) do not inhibit DCE activity indicating that DCE is distinct from these two enzymes. Cleavage site determination with matrix-assisted laser desorption ionization time of flight (MALDITOF) mass spectrometry shows that DCE cleaves the Dyn B-29 N terminus to the Arg(14) generating Dyn B-13 and Dyn B-(14-29). Among other peptides derived from Dyn B-29, DCE cleaves only those peptides that fit the predicted ''consensus motif'' for monobasic processing. These data are consistent with a broader role for the dynorphin converting enzyme in the biosynthesis of many peptide hormones and neuropeptides by processing at monobasic sites.NYU,MED CTR,DEPT PHARMACOL,NEW YORK,NY 10016ESCOLA PAULISTA MED,BR-04044020 SAO PAULO,BRAZILESCOLA PAULISTA MED,BR-04044020 SAO PAULO,BRAZILWeb of Science23845-23850engAmer Soc Biochemistry Molecular Biology IncJournal Of Biological ChemistryPURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASEinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP11600/442032021-10-05 21:55:28.38metadata only accessoai:repositorio.unifesp.br:11600/44203Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-05-25T12:27:34.036372Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
PURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASE |
title |
PURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASE |
spellingShingle |
PURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASE Berman, Y. L. |
title_short |
PURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASE |
title_full |
PURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASE |
title_fullStr |
PURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASE |
title_full_unstemmed |
PURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASE |
title_sort |
PURIFICATION AND CHARACTERIZATION OF A DYNORPHIN-PROCESSING ENDOPEPTIDASE |
author |
Berman, Y. L. |
author_facet |
Berman, Y. L. Juliano, Luiz [UNIFESP] Devi, L. A. |
author_role |
author |
author2 |
Juliano, Luiz [UNIFESP] Devi, L. A. |
author2_role |
author author |
dc.contributor.institution.none.fl_str_mv |
NYU Universidade Federal de São Paulo (UNIFESP) |
dc.contributor.author.fl_str_mv |
Berman, Y. L. Juliano, Luiz [UNIFESP] Devi, L. A. |
description |
Dynorphin B (Dyn B-13, also known as rimorphin) is generated from Dyn B-29 (leumorphin) by the cleavage at a single Arg residue. An enzymatic activity capable of processing at this monobasic site has been previously reported in neurosecretory vesicles of the bovine pituitary and pituitary-derived cell lines. This enzyme termed ''the dynorphin-converting enzyme'' (DCE) has been purified to apparent homogeneity from the neurointermediate lobe of the bovine pituitary using hydrophobic chromatography on phenyl-Sepharose, preparative isoelectrofocusing in a granulated gel between pH 4 to 6.5, and non-denaturing electrophoresis on 5% polyacrylamide gel, DCE exhibits a pI of about 5.1 and a molecular mass of about 54 kDa under reducing conditions. DCE is a metallopeptidase and exhibits a neutral pH optimum. Specific Inhibitors of soluble metallopeptidases such as enkephalinase (EC 3.4.24.11) or enkephalin generating neutral endopeptidase (EC 3.4.24.15) do not inhibit DCE activity indicating that DCE is distinct from these two enzymes. Cleavage site determination with matrix-assisted laser desorption ionization time of flight (MALDITOF) mass spectrometry shows that DCE cleaves the Dyn B-29 N terminus to the Arg(14) generating Dyn B-13 and Dyn B-(14-29). Among other peptides derived from Dyn B-29, DCE cleaves only those peptides that fit the predicted ''consensus motif'' for monobasic processing. These data are consistent with a broader role for the dynorphin converting enzyme in the biosynthesis of many peptide hormones and neuropeptides by processing at monobasic sites. |
publishDate |
1995 |
dc.date.issued.fl_str_mv |
1995-10-06 |
dc.date.accessioned.fl_str_mv |
2018-06-15T17:53:04Z |
dc.date.available.fl_str_mv |
2018-06-15T17:53:04Z |
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.citation.fl_str_mv |
Journal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 270, n. 40, p. 23845-23850, 1995. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/11600/44203 http://doi.org/10.1074/jbc.270.40.23845 |
dc.identifier.issn.none.fl_str_mv |
0021-9258 |
dc.identifier.doi.none.fl_str_mv |
10.1074/jbc.270.40.23845 |
dc.identifier.wos.none.fl_str_mv |
WOS:A1995RY90900091 |
identifier_str_mv |
Journal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 270, n. 40, p. 23845-23850, 1995. 0021-9258 10.1074/jbc.270.40.23845 WOS:A1995RY90900091 |
url |
http://repositorio.unifesp.br/11600/44203 http://doi.org/10.1074/jbc.270.40.23845 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Journal Of Biological Chemistry |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
23845-23850 |
dc.publisher.none.fl_str_mv |
Amer Soc Biochemistry Molecular Biology Inc |
publisher.none.fl_str_mv |
Amer Soc Biochemistry Molecular Biology Inc |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
|
_version_ |
1783460293229150208 |