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RSC

Atomic Resolution Structures of Mitochondrial Respiratory Chain Supercomplexes

Total Cost €

0

EC-Contrib. €

0

Partnership

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 RSC project word cloud

Explore the words cloud of the RSC project. It provides you a very rough idea of what is the project "RSC" about.

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Project "RSC" data sheet

The following table provides information about the project.

Coordinator
INSTITUTE OF SCIENCE AND TECHNOLOGYAUSTRIA 

Organization address
address: Am Campus 1
city: KLOSTERNEUBURG
postcode: 3400
website: www.ist.ac.at

contact info
title: n.a.
name: n.a.
surname: n.a.
function: n.a.
email: n.a.
telephone: n.a.
fax: n.a.

 Coordinator Country Austria [AT]
 Project website https://ist.ac.at/research/research-groups/sazanov-group/
 Total cost 178˙156 €
 EC max contribution 178˙156 € (100%)
 Programme 1. H2020-EU.1.3.2. (Nurturing excellence by means of cross-border and cross-sector mobility)
 Code Call H2020-MSCA-IF-2015
 Funding Scheme MSCA-IF-EF-ST
 Starting year 2016
 Duration (year-month-day) from 2016-09-01   to  2018-08-31

 Partnership

Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
1    INSTITUTE OF SCIENCE AND TECHNOLOGYAUSTRIA AT (KLOSTERNEUBURG) coordinator 178˙156.00

Map

 Project objective

The objectives of this proposal are to determine atomic-resolution structures of mammalian mitochondrial respiratory chain supercomplexes (RCSs) by single particle electron cryo-microscopy (cryo-EM). It has been shown that the large protein complexes of the inner mitochondrial membrane responsible for electron transport from NADH to molecular oxygen (complexes I, III and IV) associate to form three major RCSs: I/III2, III2/IV2 and I/III2/IV. Currently, the physiological role of the RSCs is controversial and poorly understood. Atomic-resolution structures of the RSCs will be essential for fully understanding their physiology. Presently I have generated a 15 Å cryo-EM density map for RCS I/III2/IV from ~10,000 particles purified from Ovis aries mitochondria. Our preliminary data show that in order to push the resolution we will need to improve both the biochemical preparation of the RCS particles as well as the cryo-EM grid preparation. This will be done through systematic biochemical characterisation of the RCSs in different non-disruptive detergents and systematic grid preparation varying grid type and blotting conditions. In addition to RCS I/III2/IV, the structure of RCS I/III2 will be determined. High-resolution cryo-EM data will be collected at the Vienna Biocenter using the FEI Polara 300 mV microscope with Gatan K2 direct electron detector. In order to fully compare the make-up of the different RCSs mass spectrometric analysis will be carried out on the purified samples by our collaborators at the Laboratory for Molecular Biology (LMB). My previous experience in the laboratory of Dr Roderick MacKinnon mainly focused on membrane protein structure determination by X-ray crystallography and NMR spectroscopy. The opportunity to work at the IST Austria with Dr Leonid Sazanov will allow me to learn state of the art cryo-EM techniques, which are positioned to be the future of atomic-resolution structure determination for large macromolecular assemblies.

 Publications

year authors and title journal last update
List of publications.
2016 James A. Letts, Karol Fiedorczuk, Leonid A. Sazanov
The architecture of respiratory supercomplexes
published pages: 644-648, ISSN: 0028-0836, DOI: 10.1038/nature19774
Nature 537/7622 2019-06-13
2016 Fiedorczuk, Karol; Degliesposti, Gianluca; Sazanov, Leonid A.; Skehel, Mark; Letts, James A
Purification of Ovine Respiratory Complex I Results in a Highly Active and Stable Preparation
published pages: , ISSN: 1083-351X, DOI: 10.1074/jbc.M116.735142
Journal of Biological Chemistry 1 2019-06-13

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