Specificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acids
Autor(a) principal: | |
---|---|
Data de Publicação: | 1999 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/26065 http://dx.doi.org/10.1042/0264-6021:3390473 |
Resumo: | We have explored in detail the determinants of specificity for the hydrolysis by human tissue kallikrein (hK(1)) of substrates containing the Phe-Phe amino acid pair, after which hK(1) cleaves kallistatin (human kallikrein-binding protein), a specific serpin for this protease, as well as somatostatin 1-14. Internally quenched fluorogenic peptides were synthesized with the general structure Abz-peptidyl-EDDnp [Abz, o-aminobenzoic acid; EDDnp, N-(2,4-dinitrophenyl)ethylenediamine], based on the natural reactive-centre loop sequence of kallistatin from P-9 to P'(13), and the kinetic parameters of their hydrolysis by hK(1) were determined. All these peptides were cleaved after the Phe-Phe pair. for comparison, we have also examined peptides containing the reactive-centre loop sequences of human protein-C inhibitor (PCI) and rat kallikrein-binding protein, which were hydrolysed after Phe-Arg and Leu-Lys bonds, respectively. Hybrid peptides containing kallistatin-PCI sequences showed that the efficiency of hK(1) activity on the peptides containing kallistatin and PCI sequences depended on both the nature of the P-1 amino acid as well as on residues at the P- and PI-sides. Moreover, we have made systematic modifications on the hydrophobic pair Phe-Phe, and on Lys and lie at the P-3 and P-4 positions according to the peptide substrate, Abz-AIKFFSRQ-EDDnp, All together, we concluded that tissue kallikrein was very effective on short substrates that are cleaved after the Phe-Arg pair; however, hydrolysis after Phe-Phe or other hydrophobic pairs of amino acids was more restrictive, requiring additional enzyme-substrate interaction and/or particular substrate conformations. |
id |
UFSP_097af456fb314217c49a32bba1752666 |
---|---|
oai_identifier_str |
oai:repositorio.unifesp.br:11600/26065 |
network_acronym_str |
UFSP |
network_name_str |
Repositório Institucional da UNIFESP |
repository_id_str |
3465 |
spelling |
Pimenta, Daniel C.Chao, JulieChao, LeeJuliano, Maria Aparecida [UNIFESP]Juliano, Luiz [UNIFESP]Universidade Federal de São Paulo (UNIFESP)Med Univ S Carolina2016-01-24T12:30:48Z2016-01-24T12:30:48Z1999-04-15Biochemical Journal. London: Portland Press, v. 339, p. 473-479, 1999.0264-6021http://repositorio.unifesp.br/handle/11600/26065http://dx.doi.org/10.1042/0264-6021:339047310.1042/0264-6021:3390473WOS:000079918400033We have explored in detail the determinants of specificity for the hydrolysis by human tissue kallikrein (hK(1)) of substrates containing the Phe-Phe amino acid pair, after which hK(1) cleaves kallistatin (human kallikrein-binding protein), a specific serpin for this protease, as well as somatostatin 1-14. Internally quenched fluorogenic peptides were synthesized with the general structure Abz-peptidyl-EDDnp [Abz, o-aminobenzoic acid; EDDnp, N-(2,4-dinitrophenyl)ethylenediamine], based on the natural reactive-centre loop sequence of kallistatin from P-9 to P'(13), and the kinetic parameters of their hydrolysis by hK(1) were determined. All these peptides were cleaved after the Phe-Phe pair. for comparison, we have also examined peptides containing the reactive-centre loop sequences of human protein-C inhibitor (PCI) and rat kallikrein-binding protein, which were hydrolysed after Phe-Arg and Leu-Lys bonds, respectively. Hybrid peptides containing kallistatin-PCI sequences showed that the efficiency of hK(1) activity on the peptides containing kallistatin and PCI sequences depended on both the nature of the P-1 amino acid as well as on residues at the P- and PI-sides. Moreover, we have made systematic modifications on the hydrophobic pair Phe-Phe, and on Lys and lie at the P-3 and P-4 positions according to the peptide substrate, Abz-AIKFFSRQ-EDDnp, All together, we concluded that tissue kallikrein was very effective on short substrates that are cleaved after the Phe-Arg pair; however, hydrolysis after Phe-Phe or other hydrophobic pairs of amino acids was more restrictive, requiring additional enzyme-substrate interaction and/or particular substrate conformations.UNIFESP, Escola Paulista Med, Dept Biofis, BR-04044020 São Paulo, BrazilMed Univ S Carolina, Dept Biochem & Mol Biol, Charleston, SC 29425 USAUNIFESP, Escola Paulista Med, Dept Biofis, BR-04044020 São Paulo, BrazilWeb of Science473-479engPortland PressBiochemical Journalfluorescent substratekallistatinpeptideproteaseserpinSpecificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acidsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP11600/260652021-09-29 11:23:43.868metadata only accessoai:repositorio.unifesp.br:11600/26065Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652021-09-29T14:23:43Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Specificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acids |
title |
Specificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acids |
spellingShingle |
Specificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acids Pimenta, Daniel C. fluorescent substrate kallistatin peptide protease serpin |
title_short |
Specificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acids |
title_full |
Specificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acids |
title_fullStr |
Specificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acids |
title_full_unstemmed |
Specificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acids |
title_sort |
Specificity of human tissue kallikrein towards substrates containing Phe-Phe pair of amino acids |
author |
Pimenta, Daniel C. |
author_facet |
Pimenta, Daniel C. Chao, Julie Chao, Lee Juliano, Maria Aparecida [UNIFESP] Juliano, Luiz [UNIFESP] |
author_role |
author |
author2 |
Chao, Julie Chao, Lee Juliano, Maria Aparecida [UNIFESP] Juliano, Luiz [UNIFESP] |
author2_role |
author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) Med Univ S Carolina |
dc.contributor.author.fl_str_mv |
Pimenta, Daniel C. Chao, Julie Chao, Lee Juliano, Maria Aparecida [UNIFESP] Juliano, Luiz [UNIFESP] |
dc.subject.eng.fl_str_mv |
fluorescent substrate kallistatin peptide protease serpin |
topic |
fluorescent substrate kallistatin peptide protease serpin |
description |
We have explored in detail the determinants of specificity for the hydrolysis by human tissue kallikrein (hK(1)) of substrates containing the Phe-Phe amino acid pair, after which hK(1) cleaves kallistatin (human kallikrein-binding protein), a specific serpin for this protease, as well as somatostatin 1-14. Internally quenched fluorogenic peptides were synthesized with the general structure Abz-peptidyl-EDDnp [Abz, o-aminobenzoic acid; EDDnp, N-(2,4-dinitrophenyl)ethylenediamine], based on the natural reactive-centre loop sequence of kallistatin from P-9 to P'(13), and the kinetic parameters of their hydrolysis by hK(1) were determined. All these peptides were cleaved after the Phe-Phe pair. for comparison, we have also examined peptides containing the reactive-centre loop sequences of human protein-C inhibitor (PCI) and rat kallikrein-binding protein, which were hydrolysed after Phe-Arg and Leu-Lys bonds, respectively. Hybrid peptides containing kallistatin-PCI sequences showed that the efficiency of hK(1) activity on the peptides containing kallistatin and PCI sequences depended on both the nature of the P-1 amino acid as well as on residues at the P- and PI-sides. Moreover, we have made systematic modifications on the hydrophobic pair Phe-Phe, and on Lys and lie at the P-3 and P-4 positions according to the peptide substrate, Abz-AIKFFSRQ-EDDnp, All together, we concluded that tissue kallikrein was very effective on short substrates that are cleaved after the Phe-Arg pair; however, hydrolysis after Phe-Phe or other hydrophobic pairs of amino acids was more restrictive, requiring additional enzyme-substrate interaction and/or particular substrate conformations. |
publishDate |
1999 |
dc.date.issued.fl_str_mv |
1999-04-15 |
dc.date.accessioned.fl_str_mv |
2016-01-24T12:30:48Z |
dc.date.available.fl_str_mv |
2016-01-24T12:30:48Z |
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. 339, p. 473-479, 1999. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/26065 http://dx.doi.org/10.1042/0264-6021:3390473 |
dc.identifier.issn.none.fl_str_mv |
0264-6021 |
dc.identifier.doi.none.fl_str_mv |
10.1042/0264-6021:3390473 |
dc.identifier.wos.none.fl_str_mv |
WOS:000079918400033 |
identifier_str_mv |
Biochemical Journal. London: Portland Press, v. 339, p. 473-479, 1999. 0264-6021 10.1042/0264-6021:3390473 WOS:000079918400033 |
url |
http://repositorio.unifesp.br/handle/11600/26065 http://dx.doi.org/10.1042/0264-6021:3390473 |
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 |
473-479 |
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_ |
1802764143405563904 |