Work

Measuring Exocytosis Rate Using Corrected Fluorescence Recovery After Photoconversion

Year: 2016

Type: article

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 changes in the photoconverted red signal are measured, and the rate of exocytosis is calculated from the changing rates of these two signals.

Source: Traffic

DOI: 10.1111/tra.12380

Authors: Nan Luo, An Yan, Zhenbiao Yang

Institutions: University of California, Riverside, Institute for Integrative Genome Biology, Center for Plant Cell Biology, California Institute of Technology

Language: English

Topic: Cellular transport and secretion

Subfield: Cell Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funder: National Institute of General Medical Sciences

Awards: Mechanical Regulation of Apical and Invasive Growth, G6910934270