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Interaction Studies of Secreted Aspartic Proteases (Saps) from Candida albicans: Application for Drug Discovery
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry.
2005 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis is focused on enzymatic studies of the secreted aspartic proteases (Saps) from Candida albicans as a tool for discovery of anti-candida drugs. C. albicans causes infections in a number of different locations, which differ widely in the protein substrates available and pH. Since C. albicans needs Saps during virulent growth, these enzymes are good targets for drug development.

In order to investigate the catalytic characteristics of Saps and their inhibitor affinities, substrate-based kinetic assays were developed. Due to the low sensitivity of these assays, especially at the sub-optimal pH required to mimic the different locations of infections, these assays were not satisfactory. Therefore, a biosensor assay was developed whereby, it was possible to study interaction between Saps and inhibitors without the need to optimise catalytic efficacy. Furthermore, the biosensor assay allowed determination of affinity, as well as the individual association and dissociation rates for inhibitor interactions.

Knowledge about substrate specificity, Sap subsite adaptivity, and the pH dependencies of catalytic efficacy has been accumulated. Also, screening of transition-state analogue inhibitors designed for HIV-1 protease has revealed inhibitors with affinity for Saps. Furthermore, the kinetics and pH dependencies of their interaction with Saps have been investigated. One of these inhibitors, BEA-440, displayed a complex interaction with Saps, indicating a conformational change upon binding and a very slow dissociation rate. A time dependent interaction was further supported by inhibition measurements. The structural information obtained affords possibilities for design of new more potent inhibitors that might ultimately become drugs against candidiasis. The strategy to combine substrate specificity studies with inhibitor screening has led to complementary results that generate a framework for further development of potent inhibitors.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis , 2005. , p. 53
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 58
Keywords [en]
Biochemistry, SPR Biosensor, Secreted aspartic proteases, Candida albicans, interaction kinetics, drug discovery, protease inhibitor
Keywords [sv]
Biokemi
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-5815ISBN: 91-554-6263-4 (print)OAI: oai:DiVA.org:uu-5815DiVA, id: diva2:166496
Public defence
2005-06-03, Room C8:305, BMC, Husargatan 3, Uppsala, 13:15
Opponent
Supervisors
Available from: 2005-05-12 Created: 2005-05-12 Last updated: 2017-05-04Bibliographically approved
List of papers
1. Discovery of the subsite specificity of Candida albicans Sap1 and Sap2 and the design of potent peptidomimetic inhibitors
Open this publication in new window or tab >>Discovery of the subsite specificity of Candida albicans Sap1 and Sap2 and the design of potent peptidomimetic inhibitors
Show others...
(English)Manuscript (Other (popular science, discussion, etc.))
Identifiers
urn:nbn:se:uu:diva-93113 (URN)
Available from: 2005-05-12 Created: 2005-05-12 Last updated: 2009-11-27Bibliographically approved
2. Substrate specificity and pH dependence of secreted aspartic proteases Sap1, Sap2 and Sap3 from Candida albicans
Open this publication in new window or tab >>Substrate specificity and pH dependence of secreted aspartic proteases Sap1, Sap2 and Sap3 from Candida albicans
(English)Manuscript (Other (popular science, discussion, etc.))
Identifiers
urn:nbn:se:uu:diva-93114 (URN)
Available from: 2005-05-12 Created: 2005-05-12 Last updated: 2009-11-27Bibliographically approved
3. Kinetic and mechanistic analysis of the association and dissociation of inhibitors interacting with secreted aspartic acid proteases 1 and 2 from Candida albicans
Open this publication in new window or tab >>Kinetic and mechanistic analysis of the association and dissociation of inhibitors interacting with secreted aspartic acid proteases 1 and 2 from Candida albicans
2003 (English)In: Biochimica et Biophysica Acta - Proteins and Proteomics, ISSN 1570-9639, E-ISSN 1878-1454, Vol. 1646, no 1-2, p. 184-195Article in journal (Refereed) Published
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-93115 (URN)10.1016/S1570-9639(03)00022-0 (DOI)
Available from: 2005-05-12 Created: 2005-05-12 Last updated: 2017-12-14Bibliographically approved
4. Biosensor-based screening and characterization of HIV-1 inhibitor interactions with Sap1, Sap2 and Sap3 from Candida albicans
Open this publication in new window or tab >>Biosensor-based screening and characterization of HIV-1 inhibitor interactions with Sap1, Sap2 and Sap3 from Candida albicans
2006 (English)In: Journal of Biomolecular Screening, ISSN 1087-0571, E-ISSN 1552-454X, Vol. 11, no 2, p. 165-175Article in journal (Refereed) Published
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-93116 (URN)10.1177/1087057105284270 (DOI)16418316 (PubMedID)
Available from: 2005-05-12 Created: 2005-05-12 Last updated: 2017-12-14Bibliographically approved

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