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Single-particle cryo-EM analysis of septin heterocomplexes

Grant number: 18/20816-9
Support type:Scholarships abroad - Research Internship - Post-doctor
Effective date (Start): January 15, 2019
Effective date (End): July 14, 2019
Field of knowledge:Biological Sciences - Biophysics
Principal Investigator:Richard Charles Garratt
Grantee:Samuel Leite Guimarães
Supervisor abroad: Wah Chiu
Home Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
Local de pesquisa : Stanford University, United States  
Associated to the scholarship:17/05665-1 - Structural studies of septin hetero-complexes by electron microscopy, BP.PD

Abstract

Septins are GTPases that interact with one another to form filaments and take part in several fundamental cellular processes such as cytokinesis and plasma membrane compartmentalization. Even though there exist a growing interest in this important cytoskeletal component, several aspects of septin polymerization, bundling and function remain unclear. In an attempt to shed light on these questions it is important to solve the 3D structures of the individual septins as well as the heterofilaments that they form. Despite considerable effort, the only structure of a septin protofilament published to date is that of the human wild-type SEPT2-SEPT6-SEPT7 hetero-filament at 4 Å resolution that was solved by X-ray crystallography in 2007. Since septins protofilaments are diverse and difficult to crystallize, an alternative feasible technique capable of achieving near-atomic resolution structures is highly desirable. Cryo-Electron Microscopy is a recent Nobel Prize-winning technique that has shown rapid growth in the last decade, and is now powerful enough to achieve sub 4 Å resolution or even better. With the collaboration of Dr. Wah Chiu, an expert in cryo-EM from Stanford University, we aim to acquire know-how in sample preparation and imaging by cryo-EM, and most importantly, experience in the processing of cryo-EM data in order to solve septin filament 3D structures aiming at a more complete understanding of their structure-function relationships. This request is timely given the recent arrival of the first Titan Krios instrument in Brazil which should be operational within the next few months.

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