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بیست و نهمین کنفرانس شیمی آلی ایران
Identifying Novel Small Organic Allosteric Modulators for Fumarate Hydratase
نویسندگان :
Hamid Reza Kalhor ( دانشگاه صنعتی شریف ) , Elahe Navaee lavasani ( دانشگاه صنعتی شریف )
کلید واژه ها :
Fumarate hydratase،Double Strand Breaks،Allosteric modulators
چکیده مقاله :
Fumarate hydratase, also known as “Fumarase” is an enzyme of the tricarboxylic acid (TCA) cycle in mitochondria which converts fumarate to L-malate. In recent years, fumarase has emerged as an important player in response to DNA double strand breaks (DSBs) in the nucleus1.The maintenance of genome integrity constitutes major challenges for all living organisms, and fumarase deficiency has been linked to several pathologies including cancers in humans. The aim of the present study, is to identify allosteric modulators leading to changes in the enzyme function, without competing with its main substrate, working specifically and in a low concentration. Using PCR-cloning, the fumarase gene (from Saccharomyces cerevisiae) was amplified and cloned into a bacterial expression vector. The expression and purification of the recombinant enzyme was confirmed using gel electrophoresis. The kinetic parameters including Km and kcat were measured for the enzyme. To search for allosteric modulators, a library of small organic molecules is screened virtually, by molecular docking. The best candidate molecules are then identified experimentally, using the recombinant fumarase.
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