S3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates
Autor(a) principal: | |
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Data de Publicação: | 2003 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/27333 http://dx.doi.org/10.1042/BJ20030438 |
Resumo: | We have systematically examined the S3 to S3' subsite substrate specificity requirements of cathepsin K using internally quenched fluorescent peptides derived from the lead sequence Abz-KLRFSKQ-EDDnp [where Abz is o-aminobenzoic acid and EDDnp is N-(2,4-dinitrophenyl)ethylenediamine]. We assayed six series of peptides, in which each position except Gln was substituted with various natural amino acids. the results indicated that the S3-S1 subsite requirements are more restricted than those of S1'-S3'. Cathepsin K preferentially accommodates hydrophobic amino acids with aliphatic side chains (Leu, Ile and Val) in the S2 site. Modifications at P1 residues also have a large influence on cathepsin K activity. Positively charged residues (Arg and Lys) represent the best accepted amino acids in this position, although a particular preference for Gly was found as well. Subsite S3 accepted preferentially basic amino acids such as Lys and Arg. A broad range of amino acids was accommodated in the remaining subsites. We further explored the acceptance of a Pro residue in the P2 position by cathepsin K in order to develop specific substrates for the enzyme. Two series of peptides with the general sequences Abz-KXPGSKQ-EDDnp and Abz-KPXGSKQ-EDDnp (where X denotes the position of the amino acid that is altered) were synthesized. the substrates Abz-KPRGSKQ-EDDnp and Abz-KKPGSKQ-EDDnp were cleaved by cathepsin K at the Arg-Gly and Gly-Ser bonds respectively, and have been shown to be specific for cathepsin K when compared with other lysosomal cysteine proteases such as cathepsins L and B and with the aspartyl protease cathepsin D. |
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Repositório Institucional da UNIFESP |
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Alves, Marcio FMPuzer, LucianoCotrin, Simone Silva [UNIFESP]Juliano, Maria Aparecida [UNIFESP]Juliano, Luiz [UNIFESP]Bromme, DieterCarmona, Adriana Karaoglanovic [UNIFESP]Universidade Federal de São Paulo (UNIFESP)CUNY Mt Sinai Sch Med2016-01-24T12:33:57Z2016-01-24T12:33:57Z2003-08-01Biochemical Journal. London: Portland Press, v. 373, p. 981-986, 2003.0264-6021http://repositorio.unifesp.br/handle/11600/27333http://dx.doi.org/10.1042/BJ2003043810.1042/BJ20030438WOS:000184800300037We have systematically examined the S3 to S3' subsite substrate specificity requirements of cathepsin K using internally quenched fluorescent peptides derived from the lead sequence Abz-KLRFSKQ-EDDnp [where Abz is o-aminobenzoic acid and EDDnp is N-(2,4-dinitrophenyl)ethylenediamine]. We assayed six series of peptides, in which each position except Gln was substituted with various natural amino acids. the results indicated that the S3-S1 subsite requirements are more restricted than those of S1'-S3'. Cathepsin K preferentially accommodates hydrophobic amino acids with aliphatic side chains (Leu, Ile and Val) in the S2 site. Modifications at P1 residues also have a large influence on cathepsin K activity. Positively charged residues (Arg and Lys) represent the best accepted amino acids in this position, although a particular preference for Gly was found as well. Subsite S3 accepted preferentially basic amino acids such as Lys and Arg. A broad range of amino acids was accommodated in the remaining subsites. We further explored the acceptance of a Pro residue in the P2 position by cathepsin K in order to develop specific substrates for the enzyme. Two series of peptides with the general sequences Abz-KXPGSKQ-EDDnp and Abz-KPXGSKQ-EDDnp (where X denotes the position of the amino acid that is altered) were synthesized. the substrates Abz-KPRGSKQ-EDDnp and Abz-KKPGSKQ-EDDnp were cleaved by cathepsin K at the Arg-Gly and Gly-Ser bonds respectively, and have been shown to be specific for cathepsin K when compared with other lysosomal cysteine proteases such as cathepsins L and B and with the aspartyl protease cathepsin D.UNIFESP, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, BrazilCUNY Mt Sinai Sch Med, Dept Human Genet, New York, NY 10029 USAUNIFESP, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, BrazilWeb of Science981-986engPortland PressBiochemical Journallysosomal proteinasecysteine proteasefluorogenic substrateS3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substratesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP11600/273332021-09-29 11:24:45.237metadata only accessoai:repositorio.unifesp.br:11600/27333Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-05-25T12:16:14.329397Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
S3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates |
title |
S3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates |
spellingShingle |
S3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates Alves, Marcio FM lysosomal proteinase cysteine protease fluorogenic substrate |
title_short |
S3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates |
title_full |
S3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates |
title_fullStr |
S3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates |
title_full_unstemmed |
S3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates |
title_sort |
S3 to S3 ' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates |
author |
Alves, Marcio FM |
author_facet |
Alves, Marcio FM Puzer, Luciano Cotrin, Simone Silva [UNIFESP] Juliano, Maria Aparecida [UNIFESP] Juliano, Luiz [UNIFESP] Bromme, Dieter Carmona, Adriana Karaoglanovic [UNIFESP] |
author_role |
author |
author2 |
Puzer, Luciano Cotrin, Simone Silva [UNIFESP] Juliano, Maria Aparecida [UNIFESP] Juliano, Luiz [UNIFESP] Bromme, Dieter Carmona, Adriana Karaoglanovic [UNIFESP] |
author2_role |
author author author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) CUNY Mt Sinai Sch Med |
dc.contributor.author.fl_str_mv |
Alves, Marcio FM Puzer, Luciano Cotrin, Simone Silva [UNIFESP] Juliano, Maria Aparecida [UNIFESP] Juliano, Luiz [UNIFESP] Bromme, Dieter Carmona, Adriana Karaoglanovic [UNIFESP] |
dc.subject.eng.fl_str_mv |
lysosomal proteinase cysteine protease fluorogenic substrate |
topic |
lysosomal proteinase cysteine protease fluorogenic substrate |
description |
We have systematically examined the S3 to S3' subsite substrate specificity requirements of cathepsin K using internally quenched fluorescent peptides derived from the lead sequence Abz-KLRFSKQ-EDDnp [where Abz is o-aminobenzoic acid and EDDnp is N-(2,4-dinitrophenyl)ethylenediamine]. We assayed six series of peptides, in which each position except Gln was substituted with various natural amino acids. the results indicated that the S3-S1 subsite requirements are more restricted than those of S1'-S3'. Cathepsin K preferentially accommodates hydrophobic amino acids with aliphatic side chains (Leu, Ile and Val) in the S2 site. Modifications at P1 residues also have a large influence on cathepsin K activity. Positively charged residues (Arg and Lys) represent the best accepted amino acids in this position, although a particular preference for Gly was found as well. Subsite S3 accepted preferentially basic amino acids such as Lys and Arg. A broad range of amino acids was accommodated in the remaining subsites. We further explored the acceptance of a Pro residue in the P2 position by cathepsin K in order to develop specific substrates for the enzyme. Two series of peptides with the general sequences Abz-KXPGSKQ-EDDnp and Abz-KPXGSKQ-EDDnp (where X denotes the position of the amino acid that is altered) were synthesized. the substrates Abz-KPRGSKQ-EDDnp and Abz-KKPGSKQ-EDDnp were cleaved by cathepsin K at the Arg-Gly and Gly-Ser bonds respectively, and have been shown to be specific for cathepsin K when compared with other lysosomal cysteine proteases such as cathepsins L and B and with the aspartyl protease cathepsin D. |
publishDate |
2003 |
dc.date.issued.fl_str_mv |
2003-08-01 |
dc.date.accessioned.fl_str_mv |
2016-01-24T12:33:57Z |
dc.date.available.fl_str_mv |
2016-01-24T12:33:57Z |
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 |
Biochemical Journal. London: Portland Press, v. 373, p. 981-986, 2003. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/27333 http://dx.doi.org/10.1042/BJ20030438 |
dc.identifier.issn.none.fl_str_mv |
0264-6021 |
dc.identifier.doi.none.fl_str_mv |
10.1042/BJ20030438 |
dc.identifier.wos.none.fl_str_mv |
WOS:000184800300037 |
identifier_str_mv |
Biochemical Journal. London: Portland Press, v. 373, p. 981-986, 2003. 0264-6021 10.1042/BJ20030438 WOS:000184800300037 |
url |
http://repositorio.unifesp.br/handle/11600/27333 http://dx.doi.org/10.1042/BJ20030438 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Biochemical Journal |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
981-986 |
dc.publisher.none.fl_str_mv |
Portland Press |
publisher.none.fl_str_mv |
Portland Press |
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_ |
1783460270162575360 |