Determination of the crystal structure of the lectin BfL (Bauhinia forficata) and the effect of this protein, as well the protease inhibitors EcTI (Enterolobium contortisiliquum) and CrataBL (Crataeva tapia) on the NCI-60 cell line panel

• Grant number: 14/22649-1
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: February 01, 2015
• End date: May 31, 2015
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Maria Luiza Vilela Oliva
• Grantee: Mariana Cristina Cabral Silva
• Supervisor: Alexander Wlodawer
• Host Institution: Instituto Nacional de Farmacologia (INFAR). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Institution abroad: National Cancer Institute (NCI), United States
• Associated to the scholarship: 12/06366-4 - Investigation of mechanisms of action of the lectin BfL (Bauhinia forficata) in tumor cell lines: relation between structure and specificity., BP.PD

Abstract

Cancer is a disease that still leads to death in many countries, with the incidence > 90% of the mortality attributable to metastases (Valastyan & Weinberg, 2011). In Brazil, the estimates for 2014 point to the occurrence of 576,000 new cases of cancer. Breast cancer, the most frequent type of this disease in women, has very high mortality rate (INCA, 2014). The 60 cancer cell lines (NCI-60 panel) of the US National Cancer Institute provide an established tool for in vitro drug screening, the identification of mechanisms of action of drugs, and the approval of new chemotherapeutic agents (Gholami et al., 2013). Thus, we will screen the plant proteins BfL, EcTI, and CrataBL for their selective cytotoxic potential in the NCI-60 panel. BfL, a glycoprotein isolated from Bauhinia forficata seeds is a lectin that inhibits the viability of the MCF7 human breast cancer cell line. We have previously shown that BfL promotes cell death through caspase-9 inhibition, DNA fragmentation, cell cycle arrest, besides decreasing the expression of integrins ±6 and ²1 (Silva et al., 2014). EcTI is a polyspecific protease inhibitor isolated from Enterolobium contortisiliquum seeds that inhibits the adhesion, migration, and invasion of Hs746T human gastric cancer cells. EcTI leads to decrease of the expression of ²1 integrin with resulting reduction of the FAK and Src activation (de Paula et al., 2012). CrataBL, a lectin (glycoprotein) isolated from Crataeva tapia bark, inhibits bovine trypsin and factor Xa, leading to apoptosis of the DU145 and PC-3 human prostate cancer cell lines, with the release of mitochondrial cytochrome c and activation of caspase 3 in both types of cells (Ferreira et al., 2013). As our closely collaborating groups have previously determined the crystal structures of EcTI and CrataBL, we will now focus on the determination of the crystal structure of BfL, with the particular aim of using peptides identified in the protein structure for the tests in the NCI-60 panel, in order to correlate their structure and function. The knowledge of the pathways triggered by the proteins in tumor cells may result in discovering new tools for modulation of the tumor growth and progression, as well as for prevention of metastasis. This project will be also supervised by Dr. Jacek Lubkowski. (AU)

Scientific publications

(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)

LUBKOWSKI, JACEK; DURBIN, SARAH V.; SILVA, MARIANA C. C.; FARNSWORTH, DAVID; GILDERSLEEVE, JEFFREY C.; OLIVA, MARIA LUIZA V.; WLODAWER, ALEXANDER. Structural analysis and unique molecular recognition properties of a Bauhinia forficata lectin that inhibits cancer cell growth. FEBS Journal, v. 284, n. 3, p. 429-450, FEB 2017. (12/06366-4, 14/22649-1, 09/53766-5)

LUBKOWSKI, JACEK; DURBIN, SARAH V.; SILVA, MARIANA C. C.; FARNSWORTH, DAVID; GILDERSLEEVE, JEFFREY C.; OLIVA, MARIA LUIZA V.; WLODAWER, ALEXANDER. Structural analysis and unique molecular recognition properties of a Bauhinia forficata lectin that inhibits cancer cell growth. FEBS Journal, v. 284, n. 3, p. 22-pg., 2017-02-01. (12/06366-4, 14/22649-1, 09/53766-5)