Rithwik Premod Nambiar

I work with Onishchenko Lab to uncover the assembly mechanisms of multi-protein complexes. Cells produce numerous proteins, most of which not only fold into well-defined three-dimensional structures (Benschop et. al., 2010) but also assemble into distinct complexes of defined architecture and composition which are necessary to properly carry out their biological roles(Huttlin et.al., 2017). Recent advances in structure determination techniques have enabled the structural characterization of many multiprotein complexes such as the RNA polymerase (Cramer et.al., 2001), the ribosome (Carter et.al., 2000), mitochondrial OXPHOS complexes (Chaban et.al., 2014) and many others. However, much remains to be understood about the assembly and maintenance of multi-protein complexes and which factors mediate it. This knowledge gap can be partially attributed to the lack of unbiased methods to investigate these processes.My project aims to advance our understanding of protein complex biogenesis and functional maintenance by adapting two recently introduced methods based on quantitative massspectrometry - Complexome profiling (CP) (Heide et.al., 2012) and Kinetic Analysis of incorporation Ratesin Macromolecular Assemblies (KARMA) (Onishchenko et.al., 2020) - to generate first time-reloved assembly maps of two elaborate protein machines, the mitochondrial OXPHOS (WP1) and the Nuclear Pore Complex (NPC) (WP2). The project outcomes are expected to shed light on the biogenesis of these complexes and provide a generally applicable strategy to study the assembly of a wide variety of protein machines.