The Rational Design and Evaluation of CK2alpha Mutants Bearing Inhibitor-Refractory Amino Acid Substitutions
Master of Science
Dr. David Litchfield
CK2 is a ubiquitously expressed and constitutively active serine/threonine protein kinase that is implicated in many cellular functions. Previous studies have indicated that the generation of mutants that are less sensitive to inhibition can be advantageous when studying protein kinases. Importantly, studies have demonstrated that mutants of CK2 rendered less sensitive to inhibition are attainable. To extend these observations, mutants of CK2α were designed and evaluated to test their effect on the inhibition of CK2 by CX-4945 using in vitro enzymatic assays followed by the development of inducible cell lines. CX-4945 is a CK2 inhibitor that has demonstrated anti-tumor activity and has recently been extended into clinical trails phase II. It was demonstrated that a CK2α triple mutant (V66A/H160D/I174A) led to a reduction in the inhibition of CK2 by CX-4945. Generation of CK2 inhibitor-refractory mutants will provide valuable insight regarding the precise functions of CK2 as well as its inhibition.
Fess, Sam Reid, "The Rational Design and Evaluation of CK2alpha Mutants Bearing Inhibitor-Refractory Amino Acid Substitutions" (2015). Electronic Thesis and Dissertation Repository. 3448.