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RSC

Atomic Resolution Structures of Mitochondrial Respiratory Chain Supercomplexes

Total Cost €

EC-Contrib. €

Partnership

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Outcomes and
results

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.

techniques compare aries presently structure varying purified gatan protein mass ovis samples nmr respiratory transport crystallography detergents vienna macromolecular mackinnon microscope austria biocenter lmb sazanov 15 mitochondria structures iii2 particle membrane inner collected biology preparation preliminary 300 associate particles molecular collaborators atomic single spectrometric supercomplexes controversial opportunity me physiological spectroscopy em laboratory aring biochemical ist characterisation leonid poorly rcs fei iv2 complexes detector physiology form polara cryo 10 ray rscs map resolution assemblies dr mainly roderick disruptive responsible grid chain data k2 density mitochondrial nadh microscopy mv push oxygen rcss systematic shown blotting mammalian electron determined direct positioned

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 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.	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

List of publications.
year	authors and title	journal	last update
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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