Development of platforms to study transmembrane proteins - towards medical, agricultural and biotechnological benefits

• Grant number: 18/14799-4
• Support Opportunities: Regular Research Grants
• Start date: December 01, 2018
• End date: June 30, 2022
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Agreement: Durham University
• Mobility Program: SPRINT - Projetos de pesquisa - Mobilidade
• Principal Investigator: Ariel Mariano Silber
• Grantee: Ariel Mariano Silber
• Principal researcher abroad: Paul William Denny
• Institution abroad: Durham University (DU), England
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 16/06034-2 - The biological role of amino acids and their metabolites in Trypanosoma cruzi, AP.TEM

Abstract

The analysis of proteins, from functional, structural and biochemical perspectives, has moved forward apace in recent decades. For example, the Protein Data Bank (established in 1971) now contains the structures of >130,000 proteins, derived using X-ray diffraction, NMR and electron microscopy, compared to approximately 10,000 in 1999. >97% (>126,000) of these structures are soluble proteins which are relatively easy to express, purify, functionally characterise and crystallize. However, more than 1 in 4 of eukaryotic proteins are lipid-bound, transmembrane (TM) macromolecules (see Nature Protocols and Reviews Molecular Cell Biology collection, 2017). This paucity of data is attributable to several factors, most importantly the interface between two types of physicochemical environments usually has properties not easily predictable from a combination of the individual characteristics of each part of the system. Additionally, difficulties encountered in the isolation and assay of these complex, membrane-associated proteins, further impedes our understanding of these and their lipid interactions, which is currently based on only a tiny handful biophysical and structural studies in model systems. In order to make a step change in this arena and reach new levels of knowledge, there is a clear need for novel approaches and tools to be developed that can be used to define and characterise more of these complex, membraneous proteins. It is our goal to develop a highly collaborative multidisciplinary network with researchers from different areas of expertise in protein and lipid chemistry, and biochemistry. Sharing and exploiting expertise across Durham (bioassay development, membrane modelling, structural biology), USP (biochemistry, structural biology) and LNBio (biophysics, high throughput screening), this collaborative project will develop a multidisciplinary network towards the development of tools to understand the structure and function of TM proteins. Such tools will facilitate the acquisition of a deeper understanding of these under-studied functional units by allowing the probing of protein-membrane interactions and functional conformations, and complement current collaborative research between Durham and Sao Paulo (MRC GCRF "Network for Neglected Tropical Diseases"). In addition to allowing us to answer long standing questions such as - How do proteins interact with lipid membranes? How does the composition of a membrane affect function and location? - the technologies developed will also facilitate the further consideration of this large but mysterious group of proteins as drug targets, herbicide/pesticide targets, and engines of biosynthesis for industry. (AU)