CTP Synthetase in NDP Kinase Mutants
Presented by: Keith Nylin
Mentor: Christopher K. Mathews
Dept. of Biochemistry and Biophysics, Oregon State University

Abstract
When nucleoside diphosphokinase (NDP kinase), the enzyme that catalyzes the reaction from nucleoside diphosphates (NDP) to nucleoside triphosphates (NTP), is knocked out of an Escherichia coli cell, the pools of CTP increase 3.2 fold. This relationship strongly suggests allosteric regulation between NDP kinase and CTP synthetase, the enzyme that catalyzes the formation of CTP from UTP. Are CTP pools increasing because of an increase of CTP synthetase concentration, or increased CTP synthetase enzyme activity? To answer this question a purified sample of CTP synthetase is needed to inject into a rabbit to form antibodies. These antibodies will be used to detect the presence of CTP synthetase in a cell wear NDP kinase is knocked out, and a cefl wear NDP kinase is functional. The comparison between the two levels of CTP synthetase will provide evidence to whether the activity level or concentration level of CTP synthetase has increased.