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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.

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

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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