Microspectroscopic imaging tracks the intracellular processing of a signal transduction protein: Fluorescent-labeled protein kinase C β I

摘要

We have devised a microspectroscopic strat- egy for assessing the intracellular (re)distribution and the integrity of the primary structure of proteins involved in signal transduction. The purified proteins are fluorescent- labeled in vitro and reintroduced into the living cell. The localization and molecular state of fluorescent-labeled protein kinase C βI isozyme were assessed by a combination of quantitative confocal laser scanning microscopy, fluorescence lifetime imaging microscopy, and novel determinations of fluorescence resonance energy transfer based on photobleach- ing digital imaging microscopy.