Nicole A Fernandez
An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1
Nicole Fernandez, Connie Jaqueline Miranda, Nader Kamel, Daniel Turner, Del Benzenhafer, Susan N Bolch, Jacob T Andring, Robert McKenna, W Clay Smith
Wesley Clay Smith
College of Medicine
Arrestin-1 is involved in various protein interactions that affect signaling cascades. This poster will focus specifically on the validation of the model of the arrestin-1 and the glycolysis enzyme enolase-1 interaction. Based on this model, seven charge pair binding sites were identified to have the closest interactions with arrestin-1. The interaction between these two proteins were analyzed by performing seven targeted charge pair substitutions through site-directed mutagenesis on enolase-1 and arrestin-1 protein (this poster focusing primarily on enolase-1). After these seven mutations were put together into a single enolase-1 protein, it was expressed into Yeast GS115 cells and then purified. Using a pulldown protein assay, we were able to analyze and compare the binding between the mutated enolase-1 and the wild type arrestin-1 protein. Our findings show that the seven charge pair substitutions on enolase-1 completely blocked the binding with arrestin-1, thereby verifying the arrestin-1 and enolase-1 model.
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