Designing protease sensors for real-time imaging of trypsin activation in pancreatic cancer cells.

ABSTRACT: Acute pancreatitis is a serious and potentially fatal disease caused by intracellular trypsinogen activation. Although protease detection has been greatly facilitated by the development of protease probes capable of monitoring protease activation and inhibition, real-time quantitative measurement of protease activity in living cells remains a challenge, and the identification of the cellular compartment for trypsinogen activation is inconclusive. Here we report a novel strategy for developing trypsin sensors by grafting an enzymatic cleavage site into a sensitive location for optical change of chromophore in a single enhanced green fluorescent protein (EGFP). Our designed trypsin sensor exhibits rapid kinetic responses for protease activation and inhibition with a large ratiometric optical

SUBMITTER: Chen N 

PROVIDER: S-EPMC2739378 | biostudies-literature | 2009 Apr

REPOSITORIES: biostudies-literature