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ANALYSIS OF ANIONS AND PROTEINS RELATED TO OXALATE METABOLISM IN HUMAN ERYTHROCYTES

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abstract
The metabolism of oxalate and its related anions in human erythrocytes have not been previously studied in detail. In this study, we identified the presence and investigated the functional role of glyoxylate reductase/hydroxypyruvate reductase GR/HPR), a key protein in the metabolism of oxalate, in human erythrocytes. In addition, the presence of anions associated with oxalate metabolism was investigated using a novel technique that couples ion chromatography and mass spectrometry to reveal anions that previously eluded scientists due to their co-elution. Erythrocyte incubations with glycolate, glyoxylate, and C-13 glycolate were used to study whether these acids were precursors of oxalate or phosphoglycolate. This study also assessed a more costeffective and less-invasive diagnostic assay called Dried Blood Spot Filter Paper Assay the detection of Primary Hyperoxaluria Type 2 (PH2), a disease characterized by the dysfunction of GR/HPR. GR/HPR activity was detected in erythrocytes at levels similar other blood components such as blood mononuclear cells. Anions associated with HPR and oxalate metabolism were detected and quantified in 30 normal subjects. Erythrocyte incubations with glyoxylate revealed that elevated intracellular levels of glyoxylate produced elevated intracellular levels of oxalate. Erythrocyte incubations with glycolate showed that there was no significant increase in intracellular oxalate or phosphoglycolate levels as intracellular glycolate levels increased. The analysis of a follow-up study using C-13 glycolate erythrocyte incubations was unable to reveal the metabolic fate of glycolate in erythrocytes. The assessment of the Dried Blood Spot Filter Paper Assay identified the assay as a promising tool for the diagnosis of PH2. In conclusion, this study investigated a previously un-described metabolic pathway in human erythrocytes. Further investigation of this pathway could lead to a greater understanding of oxalate metabolism, better diagnostic assays, and superior treatment strategies for PH2.
subject
Oxalate
Primary Hyperoxaluria
Erythrocytes
Anions
contributor
Kiger, Jennifer (author)
Dawson, Paul (committee chair)
Holmes, Ross (committee member)
Lively, Mark (committee member)
Lowther, W. Todd (committee member)
Christ, George (committee member)
date
2009-06-04T20:36:20Z (accessioned)
2010-06-18T18:57:06Z (accessioned)
2009-06-04T20:36:20Z (available)
2010-06-18T18:57:06Z (available)
2009-06-04T20:36:20Z (issued)
degree
Molecular Genetics (discipline)
identifier
http://hdl.handle.net/10339/14665 (uri)
language
en_US (iso)
publisher
Wake Forest University
rights
Release the entire work immediately for access worldwide. (accessRights)
title
ANALYSIS OF ANIONS AND PROTEINS RELATED TO OXALATE METABOLISM IN HUMAN ERYTHROCYTES
type
Thesis

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