Characterization of Full-Length and Headless Isoforms of Myosin-X (2013)
Undergraduate: Sarbajeet Nagdas
Faculty Advisor: Richard Cheney
Department: Chemistry
Myosin-X (Myo10) is a molecular motor myosin that is best known for its induction and localization to the tips of filopodia- finger like cellular extensions. There is a ~237-kDa full-length Myo10 isoform that is ubiquitously expressed composed of a ¿head¿ that functions as an actin-based motor and a ¿tail¿ which contains many domains with the potential to bind to several biologically relevant molecules. In addition, a shorter ~165-kDa Myo10 isoform has also been detected that lacks most of the head domain. In order to better characterize the shorter ¿headless¿ isoform, our first aim is to confirm its expression pattern and start site. Using immunoblots, we confirmed that headless Myo10 is expressed in nervous system tissues and cell lines. We immunoprecipitated and sequenced headless Myo10 (as well as full-length Myo10) allowing us to identify headless Myo10¿s initiation site. Furthermore, the many domains in the tail in addition to other encouraging data suggest that Myo10 has interacting partners with important biological consequences. Our second aim is to co-immunoprecipitate a FLAG-tagged fusion protein of the full-length Myo10 in order to identify potential interacting partners. With a better understanding of the headless Myo10 and increasing knowledge of Myo10¿s interacting partners, we hope to elucidate Myo10¿s importance in a host of cellular processes.