Studies of GPCR and G-protein localization in membrane domains using single-molecule imaging

Provider

Ministry of Education, Youth and Sports

Programme

INTER-EXCELLENCE

Call for proposals

SMSM2020LTC01

Main participants

Mikrobiologický ústav AV ČR, v. v. i.

Contest type

VS - Public tender

Contract ID

MSMT-21080/2020-40

Project name in Czech

Studies of GPCR and G-protein localization in membrane domains using single-molecule imaging

Annotation in Czech

The main goal of the project is to determine spatial localization and temporal dynamics of interactions between the components of the megaplex in live cells at low, presumably close to physiological, expression levels. We intend to ascertain whether this complex is formed in the plasma membrane of live cells and whether GPCRs suggested to form the megaplex can specifically recruit their cognate G proteins to endosomes through clathrin-mediated endocytosis. The project objectives are: 1) Ascertain whether 2AR-V2R(Ct) and V2R efficiently recruit the Gs protein and -arrestin 1 into CCSs 2) Determine whether 2AR-V2R(Ct) and V2R are capable of binding the Gs protein and -arrestin 1 simultaneously at the plasma membrane of live cells 3) Determine the abundance and lifetimes of complexes between 2AR-V2R(Ct) or V2R, the Gs protein and/or -arrestin 1.

R&D category

ZV - Basic research

OECD FORD - main branch

30105 - Physiology (including cytology)

OECD FORD - secondary branch

10610 - Biophysics

OECD FORD - another secondary branch

30104 - Pharmacology and pharmacy

CEP - equivalent branches <br>(according to the <a href="http://www.vyzkum.cz/storage/att/E6EF7938F0E854BAE520AC119FB22E8D/Prevodnik_oboru_Frascati.pdf">converter</a>)

BO - Biophysics<br>ED - Physiology<br>FR - Pharmacology and apothecary chemistry

Provider evaluation

V - Vynikající výsledky projektu (s mezinárodním významem atd.)

Project results evaluation

We achieved all goals of the project including creation of biosensor constructs, development of advanced analysis algorithms, and determination of structural and functional basis of the non-canonical interactions between G protein-coupled receptors and G proteins. We found that the orphan receptor GPR19, unlike vasopressin receptor, can induce G protein localization into endogenous compartments.

Realization period - beginning

Jun 1, 2020

Realization period - end

Mar 10, 2023

Project status

U - Finished project

Latest support payment

Feb 17, 2023

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Data delivery code

CEP24-MSM-LT-U

Data delivery date

Jul 1, 2024

Total approved costs

3,987 thou. CZK

Public financial support

3,987 thou. CZK

Other public sources

0 thou. CZK

Non public and foreign sources

0 thou. CZK