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Combining X-Ray Crystallography with Small Angle X-Ray Scattering to Model Unstructured Regions of Nsa1 from S. Cerevisiae
Published on: January 10, 2018
Purification, crystallization and preliminary X-ray analysis of adenylosuccinate synthetase from the fungal pathogen
Ross D Blundell1, Simon J Williams, Carl A Morrow
1Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia.
Abstract:
With increasingly large immunocompromised populations around the world, opportunistic fungal pathogens such as Cryptococcus neoformans are a growing cause of morbidity and mortality. To combat the paucity of antifungal compounds, new drug targets must be investigated. Adenylosuccinate synthetase is a crucial enzyme in the ATP de novo biosynthetic pathway, catalyzing the formation of adenylosuccinate from inosine monophosphate and aspartate. Although the enzyme is ubiquitous and well characterized in other kingdoms, no crystallographic studies on the fungal protein have been performed. Presented here are the expression, purification, crystallization and initial crystallographic analyses of cryptococcal adenylosuccinate synthetase. The crystals had the symmetry of space group P2(1)2(1)2(1) and diffracted to 2.2 Å resolution.
Insights
Researchers crystallized cryptococcal adenylosuccinate synthetase, an enzyme crucial for ATP synthesis. This structural study provides a foundation for developing new antifungal drugs against opportunistic fungal infections.
Area of Science:
- Biochemistry
- Structural Biology
- Mycology
Background:
- Opportunistic fungal infections, like those caused by Cryptococcus neoformans, pose a significant health threat, particularly to immunocompromised individuals.
- There is an urgent need for novel antifungal compounds due to the limited availability of effective treatments.
- Adenylosuccinate synthetase is a key enzyme in the ATP de novo biosynthesis pathway, making it a potential antifungal drug target.
Purpose of the Study:
- To investigate cryptococcal adenylosuccinate synthetase as a potential target for new antifungal therapies.
- To obtain structural information on the fungal enzyme, which is lacking in current research.
Main Methods:
- Expression and purification of cryptococcal adenylosuccinate synthetase.
- Crystallization of the purified enzyme.
- Initial crystallographic analysis of the obtained crystals.
Main Results:
- Successfully expressed and purified cryptococcal adenylosuccinate synthetase.
- Obtained crystals of the enzyme with space group P2(1)2(1)2(1).
- The crystals diffracted X-rays to a resolution of 2.2 Å.
Conclusions:
- The study presents the first crystallographic analysis of fungal adenylosuccinate synthetase.
- These findings provide a structural basis for the rational design of inhibitors targeting this enzyme.
- This work is a critical step towards developing new antifungal drugs to combat Cryptococcus neoformans infections.
