The use of inhibitors to stabilize folded conformations of enzymes
department or school: Science, Faculty of
department or school: Chemistry, Department of
data provider: DSpace
degree campus: UBCV
creator: Reid, Stephen P.
issued: 2009-11-17T23:29:53Z
issued: 2004
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: Tay Sachs and Gaucher diseases are lysosomal storage disorders that are caused by deficiencies in the lysosomal enzymes β-hexosaminidase A and glucocerebrosidase, respectively. The glycolipid substrates of deficient β-hexosaminidase A and glucocerebrosidase accumulate in lysosomal neurons, resulting in neurodegenerative disorders. The enzyme deficiencies are caused by heritable amino acid point mutations that result in unstable and potentially misfolded enzymes. The mutant enzymes, although catalytically active in many cases, are destroyed by cellular quality control mechanisms in the endoplasmic reticulum (ER). A potential therapy for adult chronic Tay Sachs disease and Gaucher disease is the use of enzyme inhibitors as chemical chaperones. To act as a chemical chaperone the enzyme-specific inhibitor must stabilize the folded conformation of the enzyme and thus prevent enzyme destruction by cellular quality control mechanisms. The enzyme/inhibitor complex can then be transferred from the E R to the lysosome where the substrate concentration is significantly high that some enzyme activity is restored. In the chronic adult disease forms, only 10 % of normal enzyme activity is required for a patient to become asymptomatic. A competitive inhibitor for β-hexosaminidase A, NAG-thiazoline, was synthesized and shown to be a potent inhibitor o f β-hexosaminidase A and the related enzyme β-hexosaminidase B, with Ki values of 270 nM and 190 nM , respectively. Chemical denaturation studies showed that NAG-thiazoline stabilizes a bacterial hexosaminidase from Streptomyces plicatus (Sp. Hex.) and human P-hexosaminidase B against guanidine hydrochloride denaturation. Thermal denaturation studies showed that NAG-thiazoline stabilizes Sp. Hex., β-hexosaminidase B, and β- hexosaminidase A against thermal denaturation. These results provide a chemical explanation for recent results from collaborator Dr. Mike Tropak (The Hospital for Sick Children, Toronto), which showed that NAG-thia…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/15209
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note: T H E U S E O F I N H I B I T O R S T O S T A B I L I Z E F O L D E D C O N F O R M A T I O N S O F E N Z Y M E S By STEPHEN P. REID B . S c , The University of British Columbia, 2000 A THESIS SUBMITTED IN P A R T I A L F U L F I L L M E N T OF T H E REQUIREMENTS OF THE D E G R E E OF M A S T E R OF SCIENCE in T H E F A C U L T Y OF G R A D U A T E STUDIES (Department of Chemistry) We accept this thesis as conforming to the required standard T H E UNIVERSITY OF BRITISH C O L U M B I A March 2004 © Stephen P. Reid, 2004 Library Authorization In presenting this thesis in partial fulfillment of the requirements for an advanced degree at the University of British Columbia, I agree that the Library shall make it freely available for reference and study. I further agree that permission for extensive copying of this thesis for scholarly purposes may be granted by the head of my department or by his or her representatives. It is understood that copying or publication of this thesis for financial gain shall not be allowed without my written permission. Name of Author (please print) Date (dd/mm/yyyy) Title of Thesis: jy U>" •••/* k.'b; fu ,•-i W ^)f>bJi^ Degree: M , Sc, Year: 3 / t o u ^ Department of CWmo+,-<•-) The University of British Columbia Vancouver, B C Canada Abstract Tay Sachs and Gaucher diseases are lysosomal storage disorders that are caused by deficiencies in the lysosomal enzymes (3-hexosaminidase A and glucocerebrosidase, respectively. The glycol ipid substrates o f deficient P-hexosaminidase A and glucocerebrosidase accumulate in lysosomal neurons, resulting in neurodegenerative disorders. The enzyme deficiencies are caused by heritable amino acid point mutations that result in unstable and potentially misfolded enzymes. The mutant enzymes, although catalytically active in many cases, are destroyed by cellular quality control mechanisms in the endoplasmic reticulum (ER) . A potential therapy for adult chronic Tay Sachs disease and Gaucher disease is the use …
has type: Thesis/Dissertation
date issued: 2004-05
is shown at: 10.14288/1.0061210
language: eng
degree discipline: Chemistry
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
scholar level: Graduate
type: Text
identifier URI: http://hdl.handle.net/2429/15209