UNIVERSITY OF MASSACHUSETTS
The goal of this project is to develop a new set of molecular reagents that specifically deliver molecular probes to the individual subunits of functioning K+ channel complexes. The development of this novel technology will enable the facile determination of the subunit composition and membrane localization of K+ channel complexes in living cells. We are targetting the ion-conducting and regulatory subunits of two K+ channels found in vestibular cells. We will derivatize two scorpion toxins such that when they bind to their cognate K+ channel complex they will covalently label the complex with a molecular probe. The labeling efficiency, specificity and utility of the derivatized scorpion toxins will be demonstrated in electrical recordings, biochemical assays and fluorescence imaging experiments.