Direct electrochemical observation of glucosidase activity in isolated single lysosomes from a living cell.

ABSTRACT: The protein activity in individual intracellular compartments in single living cells must be analyzed to obtain an understanding of protein function at subcellular locations. The current methodology for probing activity is often not resolved to the level of an individual compartment, and the results provide an extent of reaction that is averaged from a group of compartments. To address this technological limitation, a single lysosome is sorted from a living cell via electrophoresis into a nanocapillary designed to electrochemically analyze internal solution. The activity of a protein specific to lysosomes, β-glucosidase, is determined by the electrochemical quantification of hydrogen peroxide generated from the reaction with its substrate and the associated enzymes preloaded in the nanocap

SUBMITTER: Pan R 

PROVIDER: S-EPMC5910846 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature