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Deciphering structural information of Hsp90 chaperone complexed with hop and chip co-chaperones using high resolution mass spectrometry and chemical cross-link

Grant number: 15/21930-1
Support type:Scholarships abroad - Research Internship - Post-doctor
Effective date (Start): November 20, 2015
Effective date (End): December 19, 2015
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal Investigator:Carlos Henrique Inacio Ramos
Grantee:Dev Sriranganadane
Supervisor abroad: Manfredo Riccardo Quadroni
Home Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Research place: Université de Lausanne (UNIL), Switzerland  
Associated to the scholarship:14/00170-6 - Study of Hsp90 interaction with other proteins by cross-linking and investigation of metals binding and their effects, BP.PD


The Hsp90 (heat shock protein 90 kDa) plays a key role in the maturation of at least 10% of all proteins produced by the cell, which makes it important for many cellular processes such signaling, proteostasis, epigenetics, telomere maintenance, innate immunity, etc. Obviously, changes in this 'highly connected node' leads to disturbances in cell function resulting in drastic consequences for the organism. Hsp90 appears to play a central role in the pathology of several types of cancer as many kinases are clients and are involved in the development of cancer. As a matter of fact, specific inhibitors of Hsp90 and co-chaperones are under investigation in clinical trials. No human cytosolic Hsp90 had its full three-dimensional structure determined and the mechanisms by which this chaperone functions and is modulated are not yet fully known. Even the set of co-chaperones that play key role in the processes aforementioned, are not yet completly understood. The obvious therapeutic potential that Hsp90 from human, and Hsp90 from other organisms, have, make essential to unravel the forces that stabilize their structure and the mechanisms by which their functions are modulated resulting in the maturation of proteins clients. The group of Pr. Carlos Ramos have already investigated the conformational and functional aspect of Hsp90, but as mentioned before there is still the impact ofits interaction with its co-chaperonessuch as HOP, CHIP or SGT1,to elucidate. To reach this goal we propose to analyze chemically cross-linked chaperone/co-chaperones complexes using fast MS/MS scan coupled with high resolutionmass spectrometry in order to: 1) Determine the conformational aspects of Hsp90 complexed with co-chaperones, 2) Determine the conformational aspects of co-chaperones alone and complexed with Hsp90, 3) Investigate the interaction of Hsp90 with several co-chaperones using different type of cross-link (zero length cross-link and 7-11 Å cross-link). Those analyzes will be possible grace of the contribution of Dr. ManfredoQuadronigroup who possess the state-of-the-art in Mass Spectrometryarea. The objective during one month is to pre-fractionate cross-linked samples, inject them in mass spectrometry and analyze those data.