Structural determination of the kidney type glutaminase and the search for its binding partners.

• Grant number: 10/05987-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: September 01, 2010
• End date: August 31, 2012
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Sandra Martha Gomes Dias
• Grantee: Alexandre Cassago
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 09/10875-9 - Cellular and biochemical studies of the glutaminase enzyme and its relation with cancer, AP.JP

Abstract

Besides aerobic glycolysis, the increased levels in glutamine comsuption is also a hallmark exhibited in tumors and both phenomena are required in order to maintain its highly proliferative phenotype. With recent publications showing that previously known oncogenes also regulate these metabolic pathways, we anticipate that the better understanding of both structural organization and mechanisms of activation and inhibition of key-proteins underlying the nowadays called metabolic adaptation has the potential to generate high-value therapeutic options for cancer treatment in the future. Of special interest on the glutaminolysis branch of the metabolic adaptation process is the Kidney-type glutaminase (KGA). In mammals, KGA is one of the three known isozymes of the glutaminases, the other two being Glutaminase C (GAC) and LGA (Liver-type Glutaminase). They have been described as mitochondrial enzymes, with their individual prevalences being highly dependent on the tissue studied. Besides a well-know catalytic core, these enzymes are predicted to have different domains that mediate protein:protein interactions. KGA presents ankyrin repeats and, more importantly, two consensus sequences found only in nuclear receptor co-regulator proteins. Moreover, KGA is likely the enzyme of choice in quiescent cells and understanding its regulation modes and binding partners can give us clues on how the metabolic adaptation process happens, culminating with an increase on the cell`s glutaminolytic capability. The goals of this project is to determine the crystallographic structure of KGA, conduct biochemical studies to understand its regulation by phosphate and glutamate and search for binding partners by co-immunoprecipitation and pull-down assays. Further functional characterization of such complexes can be conduct provided that time will be available.

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)

CASSAGO, ALEXANDRE; FERREIRA, AMANDA P. S.; FERREIRA, IGOR M.; FORNEZARI, CAMILA; GOMES, EMERSON R. M.; GREENE, KAI SU; PEREIRA, HUMBERTO M.; GARRATT, RICHARD C.; DIAS, SANDRA M. G.; AMBROSIO, ANDRE L. B.. Mitochondrial localization and structure-based phosphate activation mechanism of Glutaminase C with implications for cancer metabolism. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v. 109, n. 4, p. 1092-1097, JAN 24 2012. (10/05003-0, 09/10875-9, 10/05987-0)

SCOTA FERREIRA, AMANDA PETRINA; CASSAGO, ALEXANDRE; GONCALVES, KALIANDRA DE ALMEIDA; DIAS, MARILIA MEIRA; ADAMOSKI, DOUGLAS; RODRIGUES ASCENCAO, CAROLLINE FERNANDA; HONORATO, RODRIGO VARGAS; DE OLIVEIRA, JULIANA FERREIRA; FERREIRA, IGOR MONTEZE; FORNEZARI, CAMILA; et al. Active Glutaminase C Self-assembles into a Supratetrameric Oligomer That Can Be Disrupted by an Allosteric Inhibitor. Journal of Biological Chemistry, v. 288, n. 39, p. 28009-28020, SEP 27 2013. (12/14298-9, 10/05003-0, 09/10875-9, 10/05987-0, 09/54067-3, 11/06654-7)