Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F18%3A10380859" target="_blank" >RIV/00216208:11310/18:10380859 - isvavai.cz</a>

Výsledek na webu

<a href="https://doi.org/10.3791/57837" target="_blank" >https://doi.org/10.3791/57837</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3791/57837" target="_blank" >10.3791/57837</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins

Popis výsledku v původním jazyce

This method provides a fast approach for the determination of plasma membrane partitioning of any fluorescently-tagged peripherally-associated protein using the profiles of fluorescence intensity across the plasma membrane. Measured fluorescence profiles are fitted by a model for membrane and cytoplasm fluorescence distribution along a line applied perpendicularly to the cell periphery. This model is constructed from the fluorescence intensity values in reference cells expressing a fluorescently-tagged marker for cytoplasm and with FM 4-64-labeled plasma membrane. The method can be applied to various cell types and organisms; however, only plasma membranes of non-neighboring cells can be evaluated. This fast microscopy-based method is suitable for experiments, where subtle and dynamic changes of plasma membrane-associated markers are expected and need to be quantified, e.g., in the analysis of mutant versions of proteins, inhibitor treatments, and signal transduction observations. The method is implemented in a multi-platform R package that is coupled with an ImageJ macro that serves as a user-friendly interface.

Název v anglickém jazyce

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins

Popis výsledku anglicky

This method provides a fast approach for the determination of plasma membrane partitioning of any fluorescently-tagged peripherally-associated protein using the profiles of fluorescence intensity across the plasma membrane. Measured fluorescence profiles are fitted by a model for membrane and cytoplasm fluorescence distribution along a line applied perpendicularly to the cell periphery. This model is constructed from the fluorescence intensity values in reference cells expressing a fluorescently-tagged marker for cytoplasm and with FM 4-64-labeled plasma membrane. The method can be applied to various cell types and organisms; however, only plasma membranes of non-neighboring cells can be evaluated. This fast microscopy-based method is suitable for experiments, where subtle and dynamic changes of plasma membrane-associated markers are expected and need to be quantified, e.g., in the analysis of mutant versions of proteins, inhibitor treatments, and signal transduction observations. The method is implemented in a multi-platform R package that is coupled with an ImageJ macro that serves as a user-friendly interface.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10611 - Plant sciences, botany

Projekt

<a href="/cs/project/LO1417" target="_blank" >LO1417: Centrum experimentální biologie rostlin UK</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2018

Kód důvěrnosti údajů

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

Název periodika

JoVE: Journal of Visualized Experiments

ISSN

1940-087X

e-ISSN

—

Svazek periodika

Neuveden

Číslo periodika v rámci svazku

136

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

6

Strana od-do

—

Kód UT WoS článku

000444752100120

EID výsledku v databázi Scopus

2-s2.0-85049866222