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Content archived on 2024-05-27

Single-molecule imaging of twin-arginine transporter assembly

Objective

The export of proteins across the cytoplasmic membrane is an essential function for all cells. In bacteria, two distinct transport mechanisms are used: The well-studied Sec pathway transports unfolded proteins via extrusion through a membrane-bound channel. In contrast, much less is known regarding the Twin Arginine Translocation (Tat) pathway responsible for the transport of folded proteins. The complexity required for this function is beyond that of a simple conformational change, and requires the orchestration of multiple copies of multiple proteins assembled into a large protein complex.
We recently developed a new form of artificial lipid bilayer that in addition to exceptional stability, and simple reconstitution of membrane proteins, is capable of single-molecule fluorescence imaging and single-channel electrical recording with gigaohm seals. Droplet Interface Bilayers (DIBs) are created by contacting aqueous droplets in a lipid/oil solution. We propose to reconstitute the minimal components of the Tat system in Droplet Interface Bilayers to create a working in vitro model of this important biological pathway. Using this method we will exploit single-molecule imaging to dissect the individual steps of Tat-driven transport, and in particular quantify the changes in stoichiometry of the assembled complex that occur during transport.

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Topic(s)

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Call for proposal

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FP7-PEOPLE-2013-IEF
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Funding Scheme

Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.

MC-IEF - Intra-European Fellowships (IEF)

Coordinator

THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
EU contribution
€ 221 606,40
Address
WELLINGTON SQUARE UNIVERSITY OFFICES
OX1 2JD Oxford
United Kingdom

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Region
South East (England) Berkshire, Buckinghamshire and Oxfordshire Oxfordshire
Activity type
Higher or Secondary Education Establishments
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Total cost

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