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Identification of membrane-bound serine proteinase matriptase as processing enzyme of insulin-like growth factor binding protein-related protein-1 (IGFBP-rP1/angiomodulin/mac25).
Ahmed, Sanjida; Jin, Xinlian; Yagi, Motoki; Yasuda, Chie; Sato, Yuichiro; Higashi, Shouichi; Lin, Chen-Yong; Dickson, Robert B; Miyazaki, Kaoru.
Afiliación
  • Ahmed S; Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University, Japan.
FEBS J ; 273(3): 615-27, 2006 Feb.
Article en En | MEDLINE | ID: mdl-16420484
Insulin-like growth factor (IGF) binding protein-related protein-1 (IGFBP-rP1) modulates cellular adhesion and growth in an IGF/insulin-dependent or independent manner. It also shows tumor-suppressive activity in vivo. We recently found that a single-chain IGFB-rP1 is proteolytically cleaved to a two-chain form by a trypsin-like, endogenous serine proteinase, changing its biological activities. In this study, we attempted to identify the IGFBP-rP1-processing enzyme. Of nine human cell lines tested, seven cell lines secreted IGFBP-rP1 at high levels, and two of them, ovarian clear cell adenocarcinoma (OVISE) and gastric carcinoma (MKN-45), highly produced the cleaved IGFBP-rP1. Serine proteinase inhibitors effectively blocked the IGFBP-rP1 cleavage in the OVISE cell culture. The conditioned medium of OVISE cells did not cleave purified IGFBP-rP1, but their membrane fraction had an IGFBP-rP1-cleaving activity. The membrane fraction contained an 80-kDa gelatinolytic enzyme, which was identified as the membrane-type serine proteinase matriptase (MT-SP1) by immunoblotting. When the membrane fraction was separated by SDS/PAGE, the IGFBP-rP1-cleaving activity comigrated with matriptase. A soluble form of matriptase purified in an inhibitor-free form efficiently cleaved IGFBP-rP1 at the same site as that found in a naturally cleaved IGFBP-rP1. Furthermore, small interfering RNAs for matriptase efficiently blocked both the matriptase expression and the cleavage of IGBP-rP1 in OVISE cells. These results demonstrate that IGFBP-rP1 is processed to the two-chain form by matriptase on the cell surface.
Asunto(s)
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Serina Endopeptidasas / Membrana Celular / Proteínas de Unión a Factor de Crecimiento Similar a la Insulina Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: FEBS J Asunto de la revista: BIOQUIMICA Año: 2006 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Serina Endopeptidasas / Membrana Celular / Proteínas de Unión a Factor de Crecimiento Similar a la Insulina Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: FEBS J Asunto de la revista: BIOQUIMICA Año: 2006 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido