Fluorescence Polarization Binding Assay for Aspergillus fumigatus Virulence Factor UDP-Galactopyranose Mutase
Joint Authors
Qi, Jun
Oppenheimer, Michelle
Sobrado, Pablo
Source
Issue
Vol. 2011, Issue 2011 (31 Dec. 2011), pp.1-9, 9 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2011-08-21
Country of Publication
Egypt
No. of Pages
9
Main Subjects
Abstract EN
Aspergillus fumigatus is an opportunistic human pathogenic fungus responsible for deadly lung infections in immunocompromised individuals.
Galactofuranose (Galf) residues are essential components of the cell wall and play an important role in A.
fumigatus virulence.
The flavoenzyme UDP-galactopyranose mutase (UGM) catalyzes the isomerization of UDP-galactopyranose to UDP-galactofuranose, the biosynthetic precursor of Galf.
Thus, inhibitors of UGM that block the biosynthesis of Galf can lead to novel chemotherapeutics for treating A.
fumigatus-related diseases.
Here, we describe the synthesis of fluorescently labeled UDP analogs and the development of a fluorescence polarization (FP) binding assay for A.
fumigatus UGM (AfUGM).
High-affinity binding to AfUGM was only obtained with the chromophore TAMRA, linked to UDP by either 2 or 6 carbons with Kd values of 2.6 ± 0.2 μM and 3.0 ± 0.7 μM, respectively.
These values were ~6 times lower than when UDP was linked to fluorescein.
The FP assay was validated against several known ligands and displayed an excellent Z′ factor (0.79 ± 0.02) and good tolerance to dimethyl sulfoxide.
American Psychological Association (APA)
Qi, Jun& Oppenheimer, Michelle& Sobrado, Pablo. 2011. Fluorescence Polarization Binding Assay for Aspergillus fumigatus Virulence Factor UDP-Galactopyranose Mutase. Enzyme Research،Vol. 2011, no. 2011, pp.1-9.
https://search.emarefa.net/detail/BIM-477651
Modern Language Association (MLA)
Qi, Jun…[et al.]. Fluorescence Polarization Binding Assay for Aspergillus fumigatus Virulence Factor UDP-Galactopyranose Mutase. Enzyme Research No. 2011 (2011), pp.1-9.
https://search.emarefa.net/detail/BIM-477651
American Medical Association (AMA)
Qi, Jun& Oppenheimer, Michelle& Sobrado, Pablo. Fluorescence Polarization Binding Assay for Aspergillus fumigatus Virulence Factor UDP-Galactopyranose Mutase. Enzyme Research. 2011. Vol. 2011, no. 2011, pp.1-9.
https://search.emarefa.net/detail/BIM-477651
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-477651