Determination of cleavage sites of recombinant ADAM-17 in human cells

• Grant number: 11/22421-2
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: February 01, 2012
• End date: April 30, 2015
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Adriana Franco Paes Leme
• Grantee: Rebeca Kawahara Sakuma
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 10/19278-0 - Study of regulation of ADAMs in oral cancer, AP.JP
• Associated scholarship(s): 13/21603-5 - Quantification of cancer biomarkers using targeted proteomic approach, BE.EP.DD

Abstract

The objective of this study is to find new targets for ADAM-17 and determines the specific cleavage sites of this metalloproteinase using Molecular Biology, proteomic and mass spectrometry approaches. The recombinant ADAM-17 containing the extracellular domains: metalloproteinase, disintegrin-like, cysteine rich and epidermal-like will be expressed in E. coli. After expression and purification, the recombinant protease will be incubated with tumorigenic and non-tumorigenic cell lines, SCC-9 and HaCaT, respectively. The cleavage and released proteins from surface will be identified by mass spectrometry. The strategies used in this study can identify new cell and matrix targets of ADAM-17 in tumorigenic and non-tumorigenic cells and elucidate the preferential sites of this metalloproteinase that can be related to its mechanism of action, as well as, can help to peptidic and inhibitor design. (AU)

Scientific publications

(7)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

KAWAHARA, REBECA; GRANATO, DANIELA CAMPOS; YOKOO, SAMI; DOMINGUES, ROMENIA RAMOS; TRINDADE, DANIEL MARAGNO; PAES LEME, ADRIANA FRANCO. Mass spectrometry-based proteomics revealed Glypican-1 as a novel ADAM17 substrate. JOURNAL OF PROTEOMICS, v. 151, n. SI, p. 53-65, JAN 16 2017. (11/22421-2, 09/54067-3, 10/19278-0)

KAWAHARA, REBECA; MEIRELLES, GABRIELA V.; HEBERLE, HENRY; DOMINGUES, ROMENIA R.; GRANATO, DANIELA C.; YOKOO, SAMI; CANEVAROLO, RAFAEL R.; WINCK, FLAVIAV.; RIBEIRO, ANA CAROLINA P.; BRANDAO, THAIS BIANCA; et al. Integrative analysis to select cancer candidate biomarkers to targeted validation. ONCOTARGET, v. 6, n. 41, p. 43635-43652, DEC 22 2015. (11/22421-2, 09/54067-3, 09/53839-2, 10/19278-0)

FLORES, ISADORA L.; KAWAHARA, REBECA; MIGUEL, MARCIA C. C.; GRANATO, DANIELA C.; DOMINGUES, ROMENIA R.; MACEDO, CAROLINA C. S.; CARNIELLI, CAROLINA M.; YOKOO, SAMI; RODRIGUES, PRISCILA C.; MONTEIRO, BARBARA V. B.; et al. EEF1D modulates proliferation and epithelial-mesenchymal transition in oral squamous cell carcinoma. Clinical Science, v. 130, n. 10, p. 785-799, MAY 1 2016. (11/22421-2, 09/53839-2, 09/54067-3, 13/01607-6, 10/19278-0)

KAWAHARA, REBECA; LIMA, RENATO NIYAMA; DOMINGUES, ROMENIA R.; PAULETTI, BIANCA ALVES; MEIRELLES, GABRIELA V.; ASSIS, MICHELLE; MIGLIORINI FIGUEIRA, ANA CAROLINA; PAES LEME, ADRIANA FRANCO. Deciphering the Role of the ADAM17-Dependent Secretome in Cell Signaling. JOURNAL OF PROTEOME RESEARCH, v. 13, n. 4, p. 2080-2093, APR 2014. (11/22421-2, 09/54067-3, 09/53839-2, 10/19278-0)

GRANATO, DANIELA C.; ZANETTI, MARIANA R.; KAWAHARA, REBECA; YOKOO, SAMI; DOMINGUES, ROMENIA R.; ARAGAO, ANNELIZE Z.; AGOSTINI, MICHELLE; CARAZZOLLE, MARCELO F.; VIDAL, RAMON O.; FLORES, ISADORA L.; et al. Integrated Proteomics Identified Up-Regulated Focal Adhesion-Mediated Proteins in Human Squamous Cell Carcinoma in an Orthotopic Murine Model. PLoS One, v. 9, n. 5, MAY 23 2014. (11/22421-2, 11/02267-9, 09/53839-2, 09/54067-3, 10/19278-0)

KAWAHARA, REBECA; GRANATO, DANIELA CAMPOS; YOKOO, SAMI; DOMINGUES, ROMENIA RAMOS; TRINDADE, DANIEL MARAGNO; PAES LEME, ADRIANA FRANCO. Mass spectrometry-based proteomics revealed Glypican-1 as a novel ADAM17 substrate. JOURNAL OF PROTEOMICS, v. 151, p. 13-pg., 2017-01-16. (09/54067-3, 11/22421-2, 10/19278-0)

KAWAHARA, REBECA; GRANATO, DANIELA C.; CARNIELLI, CAROLINA M.; CERVIGNE, NILVA K.; OLIVERIA, CARINE E.; MARTINEZ, CESAR A. R.; YOKOO, SAMI; FONSECA, FELIPE P.; LOPES, MARCIO; SANTOS-SILVA, ALAN R.; et al. Agrin and Perlecan Mediate Tumorigenic Processes in Oral Squamous Cell Carcinoma. PLoS One, v. 9, n. 12, DEC 15 2014. (11/22421-2, 09/54067-3, 10/19278-0)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

SAKUMA, Rebeca Kawahara. Using mass spectrometry-based proteomics to discover cancer candidate biomarkers and to identify novel ADAM17 substrates. 2015. Doctoral Thesis - Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia Campinas, SP.