Northern Kentucky University
Quantification of Inhibitor Studies of Na+ , K+ -ATPase and H+ , K+ -ATPase Chimera
Institution
Northern Kentucky University
Faculty Advisor/ Mentor
Diana McGill
Abstract
Intracellular ion transporters such as the Na+ , K+ -ATPase (NKA) and H+ , K+ -ATPase (HKA) can be used as targets to treat diseases. The Na+ , K+ -ATPase is a target for digoxin analogs in the treatment of congestive heart failure (CHF). While the H+ , K+ - ATPase is a target for omperazole analogs in the treatment of ulcers. Chimeras of the two proteins have been produced to allow study of the drug binding specificities of these two proteins. To determine inhibitory response to digoxin analogs, multiple clonal cell lines expressing either an unaltered NKA or an NKA-HKA chimera (DB) were assayed using ouabain as the inhibitor. Isolated membranes from the cell lines were used in enzyme inhibition assays. To date only the DB chimera data have been studied. Once assays on NKA have been completed, comparison between the two will give insight into ouabain binding sites and binding specificity. Information on the inhibitory sites of the native sodium pump could provide valuable information and possible new treatment options of CHF.
Quantification of Inhibitor Studies of Na+ , K+ -ATPase and H+ , K+ -ATPase Chimera
Intracellular ion transporters such as the Na+ , K+ -ATPase (NKA) and H+ , K+ -ATPase (HKA) can be used as targets to treat diseases. The Na+ , K+ -ATPase is a target for digoxin analogs in the treatment of congestive heart failure (CHF). While the H+ , K+ - ATPase is a target for omperazole analogs in the treatment of ulcers. Chimeras of the two proteins have been produced to allow study of the drug binding specificities of these two proteins. To determine inhibitory response to digoxin analogs, multiple clonal cell lines expressing either an unaltered NKA or an NKA-HKA chimera (DB) were assayed using ouabain as the inhibitor. Isolated membranes from the cell lines were used in enzyme inhibition assays. To date only the DB chimera data have been studied. Once assays on NKA have been completed, comparison between the two will give insight into ouabain binding sites and binding specificity. Information on the inhibitory sites of the native sodium pump could provide valuable information and possible new treatment options of CHF.