A micropatterning platform for quantifying interaction kinetics between the T cell receptor and an intracellular binding protein.

ABSTRACT: A complete understanding of signaling processes at the plasma membrane depends on a quantitative characterization of the interactions of the involved proteins. Fluorescence recovery after photobleaching (FRAP) is a widely used and convenient technique to obtain kinetic parameters on protein interactions in living cells. FRAP experiments to determine unbinding time constants for proteins at the plasma membrane, however, are often hampered by non-specific contributions to the fluorescence recovery signal. On the example of the interaction between the T cell receptor (TCR) and the Syk kinase ZAP70, we present here an approach based on protein micropatterning that allows the elimination of such non-specific contributions and considerably simplifies analysis of FRAP data. Specifically, detectio

SUBMITTER: Motsch V 

PROVIDER: S-EPMC6397226 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature