OVER-EXPRESSION OF THE YEAST ATP SYNTHASE SUBUNIT-D IN ESCHERICHIA-COLI - USE OF POLYCLONAL ANTIBODIES DIRECTED AGAINST RECOMBINANT SUBUNIT-D

The yeast ATP synthase subunit d was over-expressed in E. coli and for med inclusion bodies. It was purified by solubilization in urea and sl ow removal of the urea by stepwise dialysis in the presence of a non-i onic detergent. The resulting soluble subunit d was used to prepare po lyclonal antibodies. Blots of yeast mitochondrial proteins were probed with these antibodies. The strain disrupted in ATP4 gene encoding the subunit 4 displayed only 8% of the wild type subunit d. Antibodies ag ainst subunit d did not inhibit the wild type ATPase activity. (C) 199 4 Academic Press, Inc.