Measuring Exocytosis Rate Using Corrected Fluorescence Recovery After Photoconversion.

ABSTRACT: Exocytosis plays crucial roles in regulating the distribution and function of plasma membrane (PM) and extracellular matrix proteins. However, measuring the exocytosis rate of a specific protein by conventional methods is very difficult because of exocytosis-independent trafficking such as endocytosis, which also affects membrane protein distribution. Here, we describe a novel method, corrected fluorescence recovery after photoconversion, in which exocytosis-dependent and -independent trafficking events are measured simultaneously to accurately determine exocytosis rate. In this method, the protein-of-interest is tagged with Dendra2, a green-to-red photoconvertible fluorescent protein. Following the photoconversion of PM-localized Dendra2, both the recovery of the green signal and the chan

SUBMITTER: Luo N 

PROVIDER: S-EPMC7513930 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature