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Updated: Jun 2, 2026
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Biochemical and Structural Characterization of the Carbohydrate Transport Substrate-binding-protein SP0092
Published on: October 2, 2017
Structural and functional studies of mycobacterial IspD enzymes
Christofer Björkelid1, Terese Bergfors, Lena M Henriksson
1Department of Cell and Molecular Biology, Uppsala University, Biomedical Center, Box 596, SE-75124 Uppsala, Sweden.
Summary
Researchers targeted the IspD enzyme in the MEP pathway, crucial for pathogens like tuberculosis and malaria. Structural studies of Mycobacterium IspD provide tools for developing broad-spectrum inhibitors against these diseases.
Area of Science:
- Biochemistry
- Structural Biology
- Drug Discovery
Background:
- Pathogens causing tuberculosis and malaria utilize the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway for isopentenyl diphosphate synthesis, unlike humans who use the mevalonate pathway.
- Targeting essential microbial pathways offers a strategy for selective antimicrobial drug development.
Purpose of the Study:
- To characterize the IspD enzyme from Mycobacterium tuberculosis and Mycobacterium smegmatis for structure-based drug discovery.
- To obtain high-resolution crystal structures of IspD in complex with ligands to guide inhibitor design.
Main Methods:
- Engineering and expression of soluble enzyme constructs for Mycobacterium smegmatis and Mycobacterium tuberculosis IspD.
- X-ray crystallography to determine the structures of M. smegmatis IspD with CTP and CMP, and M. tuberculosis IspD with CTP.
- Enzymatic characterization of the M. tuberculosis IspD construct.
Main Results:
- Crystal structures of M. smegmatis IspD (with CTP and CMP) and M. tuberculosis IspD (with CTP) were determined and refined.
- Comparison of structures revealed a conserved core and potential flexibility near the active site.
- Enzymatic characterization of M. tuberculosis IspD was performed.
Conclusions:
- Well-behaved enzyme constructs and their structures are valuable tools for screening potential inhibitors.
- The conserved active site suggests that inhibitors developed against IspD could have broad-spectrum activity against MEP pathway-dependent pathogens.
