Date of Award

December 2018

Degree Type

Dissertation

Degree Name

Doctor of Philosophy

Department

Chemistry

First Advisor

Graham R Moran

Committee Members

Nicholas R Silvaggi, Arsenio A Pacheco, David N Frick, Alan W Schwabacher

Keywords

metabolite damage, metabolite repair, old yellow enzyme, pre-steady-state kinetics, renalase, transient kinetics

Abstract

The human enzyme renalase was discovered in 2005 by nephrologist Gary Desir, who claimed the enzyme is secreted by the kidney into the blood where it was said to catabolize catecholamines in order to modulate blood pressure and heart rate. It has since been shown that the enzyme is expressed in all tissues and does not react with catecholamines. The research detailed in this dissertation led to the discovery that renalase oxidizes two highly toxic isomers of NAD(P)H to form innocuous NAD(P)+. We surmised that such an important cellular function would be pervasive in nature, and our lab was the first to identify an example of a prokaryotic renalase from Pseudomonas syringae pv. phaseolicola, the first renalase identified outside of the Animalia. Crystal structures of the bacterial enzyme including that of the product- bound complex and also with NADPH (serving here as a substrate analog) bound in the active site have been solved, and its mechanism has been studied in detail using transient state kinetic methods. We contrast the specificity profile of the bacterial enzyme with that of the human enzyme, report structural features, and review the history of renalase discovery and the current state of knowledge in the literature, defining the enzyme’s recently proposed role in intracellular metabolite repair.

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