Discrimination of Kinetic Models by a Combination of Microirradiation and Fluorescence Photobleaching.

ABSTRACT: Fluorescence recovery after photobleaching (FRAP) is an excellent tool to measure the chemical rate constants of fluorescently labeled proteins in living cells. Usually FRAP experiments are conducted with the protein concentrations being in a steady state, i.e., when the association and dissociation of the proteins are equilibrated. This is a strong limitation because situations in which rate constants change with time are of great scientific interest. In this study, we present an approach in which FRAP is used shortly after DNA damage introducing laser microirradiation, which results in the recruitment of the DNA clamp protein proliferating cell nuclear antigen (PCNA) to DNA lesions. We establish different kinetic models that are compatible with the observed PCNA recruitment data if FRAP

SUBMITTER: Lengert L 

PROVIDER: S-EPMC4624168 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature