Characterization of thiol-, aspartyl-, and thiol-metallo-peptidase activities in Madin-Darby canine kidney cells

Characterization of thiol-, aspartyl-, and thiol-metallo-peptidase activities in Madin-Darby canine kidney cells

Autor Oliveira, V Google Scholar
Ferro, E. S. Google Scholar
Gomes, M. D. Google Scholar
Oshiro, MEM Google Scholar
Almeida, P. C. Google Scholar
Juliano, M. A. Google Scholar
Juliano, L. Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Universidade de São Paulo (USP)
Resumo We combined fluorogenic substrates or internally quenched fluorescent peptides with specific inhibitors in the pH profile of proleolytic activity experiments in order to detect proteolytic activities in lysates of MDCK cells. Hydrolytic activities related to cathepsin B, L, and D were observed. Serine-proteinase was not detected; however, we clearly demonstrated the presence of a thiol-metallo-endo-oligopeptidase, also called thimet-oligopeptidase (TOP). This peptidase from MDCK cells has substrate and inhibitor specificities as well as an activation profile with mercaptoethanol that are indistinguishable from the recombinant rat testis TOP (EC 3.4.24.15). in addition, polyclonal purified antibodies to this enzyme depleted the TOP activity of MDCK cells in whole homogenate. Although we present only preliminary data, TOP is secreted by MDCK cells. the presence of TOP in a phenotype polarized MDCK cells can have special significance in the cytoplasmic selection, transport, or clearance of short peptides due to restriction of the enzyme to sequences from 6 to 17 amino acids. Therefore, the MDCK cell could be a very useful cellular model with which to study some of the suggested TOP biological functions as processing of biological active peptides and antigen presentation. (C) 2000 Wiley-Liss, Inc.
Palavra-chave MDCK cells
THIMET
endooligopeptidase
cathepsin B
cathepsin L
cathepsin D
Idioma Inglês
Data de publicação 2000-01-01
Publicado em Journal of Cellular Biochemistry. New York: Wiley-liss, v. 76, n. 3, p. 478-488, 2000.
ISSN 0730-2312 (Sherpa/Romeo, fator de impacto)
Publicador Wiley-Blackwell
Extensão 478-488
Fonte http://dx.doi.org/10.1002/(SICI)1097-4644(20000301)76:3<478
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000085184500014
Endereço permanente http://repositorio.unifesp.br/handle/11600/26209

Exibir registro completo




Arquivo

Arquivo Tamanho Formato Visualização

Não existem arquivos associados a este item.

Este item está nas seguintes coleções

Buscar


Navegar

Minha conta