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hsPCF-FRET SIGNED

Real-time characterisation of neuropeptide binding to a membrane receptor involved in pain and ischemic stroke

Total Cost €

0

EC-Contrib. €

0

Partnership

0

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 hsPCF-FRET project word cloud

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

sites    aid    inhibitors    treat    ligand    protein    channels    ion    disorders    skills    sensitivity    fluorescence    add    linked    asic1a    personal    serves    versatile    emerged    house    directed    downsides    pathological    interactions    asic    physiological    resolution    dependent    patch    membranes    transduced    proteins    protocol    localized    serve    membrane    dynorphin    drug    scientific    pain    scope    microsecond    psychiatric    under    neuropeptide    brain    broad    stroke    few    neurological    clamp    electrophysiology    kinetics    fluorescent    expertise    fret    direct    big    local    ischemia    scaffolds    neuronal    assay    temporal    peptide    acidosis    drugs    affinity    sensing    anticipate    limited    modulate    unprecedented    hampered    living    acid    mutagenesis    opioids    binding    expand    site    acidification    generation    tissue    diseases    prevalent    intact    signal    ischemic    setup    synaptically    cells    neuropeptides    chronic    selective    time   

Project "hsPCF-FRET" 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 207˙312 €
 EC max contribution 207˙312 € (100%)
 Programme 1. H2020-EU.1.3.2. (Nurturing excellence by means of cross-border and cross-sector mobility)
 Code Call H2020-MSCA-IF-2018
 Funding Scheme MSCA-IF-EF-ST
 Starting year 2019
 Duration (year-month-day) from 2019-04-01   to  2021-03-31

 Partnership

Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
1    KOBENHAVNS UNIVERSITET DK (KOBENHAVN) coordinator 207˙312.00

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 Project objective

Under physiological conditions, localized acidification of brain tissue serves as neuronal signal that get synaptically transduced via acid-sensing ion channels (ASIC1a). Local acidosis has, however, also been linked to some of the most prevalent neurological disorders such as chronic pain, ischemic stroke and psychiatric diseases. ASIC1a has thus emerged as drug target with great potential, but no drugs are currently available that specifically target the channels under pathological conditions. A few known neuropeptides modulate ASIC1a and could thus serve as scaffolds for a new generation of ASIC1a-selective drugs to, for example, treat pain without the typical downsides of opioids. Advances have, however, been hampered by the limited understanding of detailed protein-peptide interactions. Thus, the aim of the proposed project is to directly characterize the binding of the neuropeptide Big Dynorphin to ASIC1a in real time. Here, I will use a unique in-house developed high-sensitivity fluorescence patch-clamp electrophysiology setup and establish a protocol for a FRET-based ligand-binding assay. Together with site-directed mutagenesis, this approach will be able to identify state-dependent binding sites and key interactions, and allow direct analysis of binding affinity and kinetics under pathological conditions; all in intact membranes and with unprecedented (microsecond) temporal resolution. This information will aid future design of ASIC inhibitors with the potential to treat chronic pain and ischemia. The technology developed for this work will also enable ligand-binding studies of other membrane proteins in living cells and with high temporal resolution and will thus be of great potential value for a broad field. The project will expand my existing electrophysiology skills and add highly versatile expertise in fluorescent measurements. I thus anticipate my project to have significant personal and scientific impact beyond the scope of this proposal.

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