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fastHDX for IDPs SIGNED

Revealing the Transient Structures of Intrinsically Disordered Proteins by Microfluidics-Enabled Hydrogen-Deuterium Exchange

Total Cost €

0

EC-Contrib. €

0

Partnership

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Project "fastHDX for IDPs" data sheet

The following table provides information about the project.

Coordinator
KOBENHAVNS UNIVERSITET 

Organization address
address: NORREGADE 10
city: KOBENHAVN
postcode: 1165
website: www.ku.dk

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 Denmark [DK]
 Total cost 200˙194 €
 EC max contribution 200˙194 € (100%)
 Programme 1. H2020-EU.1.3.2. (Nurturing excellence by means of cross-border and cross-sector mobility)
 Code Call H2020-MSCA-IF-2017
 Funding Scheme MSCA-IF-EF-ST
 Starting year 2018
 Duration (year-month-day) from 2018-04-01   to  2020-03-31

 Partnership

Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
1    KOBENHAVNS UNIVERSITET DK (KOBENHAVN) coordinator 200˙194.00

Map

 Project objective

This Fellowship proposal will design, fabricate and test novel microfluidic chips for the study of intrinsically disordered proteins (IDPs) by Hydrogen-Deuterium Exchange coupled with Mass Spectrometry (HDX-MS). The research program will use thiol-ene polymerisation to form all components of the microchip allowing fast, reproducible and cheap fabrication. The microchips will incorporate multiple microfluidics channels of varying length and spatially restricted monolith plugs to allow sub-second HDX reactions to occur on-chip. The microchips will circumvent the extensive and error-prone sample pre-treatment steps of current commercial HDX-MS methodology. I will be trained in state-of-the-art HDX-MS technology and will learn how to perform site-specific, light-activated thiol-ene click-chemistry reactions. The microfluidic chips will be used to probe the transient conformational changes of three challenging IDPs of biological and pharmaceutical relevance: a-synuclein, proNerve Growth Factor (proNGF) and Epsin1. α-synuclein is an extensively characterised model IDP, shown by numerous biophysical techniques including HDX-MS to possess some regions of transient structure. α-synuclein will be used during microchip development, providing valuable proof-of-concept and a framework for which to further optimize microchip design if needed. I will then characterise the conformational states and interactions of proNGF; the cleavable pro-element is reported to be disordered, but there is little knowledge of its cellular roles. Previously unreported structural elements of Epsin1, an IDP shown to mediate lipid membrane curvature, will also be probed. HDX-MS will be complemented by electron transfer dissociation (ETD) and ion mobility separation (IM-MS).To be added

 Publications

year authors and title journal last update
List of publications.
2018 Rasmus R. Svejdal, Eleanor R. Dickinson, Drago Sticker, Jörg P. Kutter, Kasper D. Rand
Thiol-ene Microfluidic Chip for Performing Hydrogen/Deuterium Exchange of Proteins at Subsecond Time Scales
published pages: 1309-1317, ISSN: 0003-2700, DOI: 10.1021/acs.analchem.8b03050
Analytical Chemistry 91/2 2020-03-05

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