Research

Life Sciences & Biotechnology

Title :

An integrated structural biology approach to unravel a unique fatty acid activation mechanism in eukaryotes

Area of research :

Life Sciences & Biotechnology

Focus area :

Fatty acid activation mechanism in eukaryotes

Principal Investigator :

Dr Rajan Sankarnarayanan, Scientist, CSIR-Centre for Cellular and Molecular Biology (CSIR-CCMB), Hyderabad

Timeline Start Year :

2020

Timeline End Year :

2025

Contact info :

Details

Executive Summary :

Objective: Generating loss of function mutation in Yeast to study the physiological role of Dip2 .Generating loss of function mutation and overexpression lines in Drosophila .Generating loss of function mutation in embryonic stem cells .Expression profiling to identify the stages for enhanced expression to obtain quality cDNA . Expression and purification optimization in BL21(DE3), Pichia and Sf9 expression systems . Biochemical characterization of various proteins . Identification of various phenotypes in loss of function mutants along with effects of different stresses . Complementation and Localization studies . Screening for Crystallization conditions for various constructs and complexes . Complementary structural characterization using SAXS, EM . Obtain quality preparative samples for performing proteomic studies after proximity labelling Obtain quality preparative samples for performing lipidomic studies and preliminary characterization of lipid preparations using thin layer chromatography . Separation of soluble and membrane protein extracts on gel electrophoresis Performing MS and MS/MS analysis to identify interacting proteins. Performing lipidomics in Multiple reaction monitoring (MRM)-mode and discovery- mode to identify the range of lipids.

Organizations involved