Skip to main content
European Commission logo
English English
CORDIS - EU research results
CORDIS
CORDIS Web 30th anniversary CORDIS Web 30th anniversary

Unveiling dynamics and substrate interactions of large protein complexes by NMR spectroscopy

Project description

NMR spectroscopy of large protein complexes

Nuclear magnetic resonance (NMR) spectroscopy identifies protein dynamics in solution on a broad range of timescales and at atomic resolution. However, NMR studies of large proteins remain challenging as the conventional NMR methods are confined to small proteins. The EU-funded NMRofLargeComplexes project aims to extend the NMR limits to large protein complexes by combining recent developments in site-specific labelling with novel NMR methods and spectrometers. This development will facilitate the study of functionally important dynamics and substrate interactions of large protein complexes, providing insight into the relationship between dynamics, structure and function. Current studies will focus on the eukaryotic exosome, a 370 kDa soluble, asymmetric decamer, and the 80 kDa NapA Na+/H+ antiporter, an integral membrane dimer involved in ion transport.

Objective

Dynamic properties of proteins are essential for their function. NMR can reveal protein dynamics in solution on a broad range of timescales and at atomic resolution. However, NMR studies of large proteins remain challenging since conventional NMR methods are constrained to small proteins. The aim of this project is to extend the NMR size limit to large protein complexes by combining recent developments in site-specific labeling schemes with novel NMR methods employing state-of-the-art spectrometers. This will facilitate the study of functionally essential but so far uncharacterized dynamics and substrate interactions of two large protein complexes, providing unprecedented insight into the relationship between dynamics, structure and function. Since most proteins are dynamic, extending the scope of protein complexes that are amenable to NMR study is of great interest to many areas of molecular biology. Studies will be conducted on the eukaryotic exosome, a 370 kDa soluble, asymmetric decamer involved in RNA degradation and processing, and the 80 kDa Na+/H+ antiporter NapA, an integral membrane dimer involved in ion transport. For the exosome, individual subunits will be 13C-methyl labeled; the employment of sophisticated NMR methods will then permit the detection of functionally essential dynamics and RNA-substrate binding patterns of the exosome. As NapA is a low-yield membrane protein, an inexpensive 19F-labeling approach will be employed to reveal global transport dynamics and local gating motions. Exosome production and all NMR experiments are to be conducted in Remco Spranger’s lab at the University of Regensburg, Germany. NapA will be produced during a secondment in David Drew’s lab at Stockholm University, Sweden. This newly established international collaboration will permit integration of methods and knowledge to study protein systems previously inaccessible to NMR, strengthen the profile of the applicant and foster research of the involved institutes.

Coordinator

UNIVERSITAET REGENSBURG
Net EU contribution
€ 162 806,40
Address
UNIVERSITATSSTRASSE 31
93053 Regensburg
Germany

See on map

Region
Bayern Oberpfalz Regensburg, Kreisfreie Stadt
Activity type
Higher or Secondary Education Establishments
Links
Total cost
€ 162 806,40