TEDs Site Phosphorylation Regulates Myosin I Motor Activity And Function In Fission Yeast

Abstract

rounds of mutagenesis and selection of those sensors with the desired characteristics in a high-throughput (HT) manner i.e. by FACS. However, there is currently no similar HT technology for sorting libraries of fluorescent protein biosensors. Thus, we have developed a novel microfluidic platform designed for high-throughput screening based on FRET change upon analyte binding. Using this platform, we sort a library of fluorescent protein Zn 2þ sensors, selecting for both amount of FRET response and binding affinity for Zn 2þ . The device is shown schematically below. The microfluidic platform incorporates a laser diode-bar optical trap, two-channel fluorescence excitation and detection, and bright-field imaging. This technology has numerous potential applications due to the great versatility in selection parameters and its ability to sort based upon cellular perturbations.