An in vivo structure-function analysis of the pathogenesis of triosephosphate isomerase deficiency.

Roland, Bartholomew P (2014) An in vivo structure-function analysis of the pathogenesis of triosephosphate isomerase deficiency. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

Triosephosphate isomerase (TPI) is a glycolytic enzyme that catalyzes the isomerization of dihydroxyacetone phosphate into glyceraldehyde 3-phosphate, a non-linear step in glycolysis not required for the production of pyruvate. Dysfunction within TPI elicits a disease called TPI deficiency; a severe, rare, autosomal recessive disorder characterized by neurological dysfunction, shortened longevity, and hemolytic anemia. Previous studies questioned whether this disease was caused by changes in metabolism or protein conformation. To address this, we have used a genomic engineering strategy in Drosophila to study the relationship between the structure of TPI and pathology. We have generated and analyzed novel high-resolution crystal structures of TPI mutant proteins, yielding basic insights into TPI dysfunction. Our data suggest the pathogenesis of TPI deficiency is unrelated to its general role in metabolism. Further, in vitro experiments demonstrate that a toxic Drosophila TPI allele is characterized by a defect in protein dimerization. Using our genomic engineering system, we have generated several novel TPI alleles that further support the hypothesis that a conformational change at the dimer interface is sufficient to elicit TPI deficiency. We have conclusively shown that gross TPI activity is not predictive of disease presence or severity. Finally, we have identified a synaptic defect caused by a dimer interface mutant that we propose is the source of neurological dysfunction in Drosophila and TPI Deficiency patients.

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Details

Item Type:	University of Pittsburgh ETD
Status:	Unpublished
Creators/Authors:	Creators Email Pitt Username ORCID Roland, Bartholomew P bpr21@pitt.edu BPR21
ETD Committee:	Title Member Email Address Pitt Username ORCID Committee Chair DeFranco, Donald B dod1@pitt.edu DOD1 Committee Member deGroat, William C wcd2@pitt.edu WCD2 Committee Member Hong, Yang yhong@pitt.edu YHONG Committee Member Levitan, Edwin S elevitan@pitt.edu ELEVITAN Thesis Advisor Palladino, Michael J mjp44@pitt.edu MJP44
Date:	18 April 2014
Date Type:	Publication
Defense Date:	13 December 2013
Approval Date:	18 April 2014
Submission Date:	13 April 2014
Access Restriction:	5 year -- Restrict access to University of Pittsburgh for a period of 5 years.
Number of Pages:	141
Institution:	University of Pittsburgh
Schools and Programs:	School of Medicine > Molecular Pharmacology
Degree:	PhD - Doctor of Philosophy
Thesis Type:	Doctoral Dissertation
Refereed:	Yes
Uncontrolled Keywords:	Drosophila, triosephosphate isomerase, triosephosphate isomerase deficiency
Date Deposited:	18 Apr 2014 12:01
Last Modified:	18 Apr 2019 05:15
URI:	http://d-scholarship.pitt.edu/id/eprint/21193

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• An in vivo structure-function analysis of the pathogenesis of triosephosphate isomerase deficiency. (deposited 18 Apr 2014 12:01) [Currently Displayed]

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