Interpretation of binding kinetics in fluorescence recovery after photobleaching experiments using a novel stochastic simulation strategy

摘要

Fluorescence recovery after photobleaching (FRAP) has been extensively used for monitoring the binding kinetics ofproteins with a goal to investigate the cellular processes, such as transcriptional regulation, cell membrane diffusion andsignal transduction. In this study, a new approach for the interpretation of FRAP curves is presented based on the stochasticsimulation of binding kinetics. The proposed method considers that proteins (a) randomly diffuse in a Brownian random-walk manner and (b) react with certain probability with compatible empty binding sites in a homogeneous well-stirredchemical environment. The proposed algorithm was compared with standard deterministic methods that are currently beingused for analysis of FRAP curves. Predictions of recovery times of FRAP curves and sum of residuals revealed a goodagreement. The stochastic simulation algorithm presents a firmer physical basis than its deterministic counterparts and itmight be used to successfully model probabilistic events in the cell, deciphering information in FRAP experiments thatcannot be computed using deterministic models.