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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.

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

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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