Structure, dynamics, evolution and catalysis of phosphohexomutase from Pseudomonas aeruginosa
No Thumbnail Available
Authors
Meeting name
Sponsors
Date
Journal Title
Format
Thesis
Subject
Abstract
[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] [alpha]-D phosphohexomutases are ubiquitous in the environment and participate in carbohydrate metabolism by catalyzing an intramolecular phosphoryl transfer. Biophysical and biochemical techniques have been utilized to study a phosphohexomutase from the human pathogen Pseudomonas aeruginosa, which contributes to its infectivity and virulence. The research work described herein includes use of computational analysis, mutagenesis, steady-state kinetics, hydrogen deuterium exchange by mass spectrometry (HDXMS), X-ray crystallography and small angle X-ray scattering (SAXS) to investigate important features underlying the catalytic mechanism. With mutagenesis and kinetics, a key amino acid in the active site of enzyme that is essential to function was identified. Computational analysis coupled with biochemical characterizations revealed a network of co-evolving residues at domain interface, site of conformational change. Based on HDXMS and SAXS studies, dephosphorylation of the enzyme results in a global increase in flexibility that coincides with a mandatory step of intermediate reorientation during the catalytic cycle. This study provided insight into the role of conformational plasticity in substrate binding and phosphorylation in the enzymatic reaction.
Table of Contents
DOI
PubMed ID
Degree
Ph. D.
Thesis Department
Rights
Access is limited to the campuses of the University of Missouri.