Publication Date
2015
Document Type
Thesis
Committee Members
Norma Adragna (Committee Member), Nadja Grobe (Committee Member), Peter Lauf (Advisor)
Degree Name
Master of Science (MS)
Abstract
In silico analysis predicts interaction between the Na/ K-ATPase (NKA) and Bcl-2 protein canonical motifs BH3 and BH1. Such interaction is consistent with NKA inhibition by the benzo-phenanthridine alkaloid chelerythrine (CHE), a BH3 mimetic, in human lens epithelial cells (HLEC). This report establishes proof of concept: co-immuno-precipitation and immuno-colocalization showed unequivocal and direct interaction between NKA and Bcl-2 proteins. Particularly, NKA-antibodies co-immunoprecipitated BclXl and BAK in three different epithelial cell lines, HLECs and A549 lung cancer cells, whereas anti-Bcl-2 antibodies failed to pull down NKA. The molecular mass of the BAK proteins pulled down by antibodies against NKA and BclXl appeared to be some 4 kDa larger than found in the input monomer. These findings are explained by NKA pulling down a BAK splice variant. Based on the lack of reversal precipitation it is suggested that both BclXl and BAK interact through different motifs with NKA. These novel findings support our hypothesis for a special sensor role of NKA in the Bcl-2 protein governance of cellular survival and apoptosis.
Page Count
70
Department or Program
Department of Pharmacology and Toxicology
Year Degree Awarded
2014
Copyright
Copyright 2015, all rights reserved. My ETD will be available under the "Fair Use" terms of copyright law. This may be required by third-party publishers you work with to publish your paper commercially.
