Molecular cloning, overexpression, purification, crystallization and preliminary X-ray diffraction studies of histidinol phosphate aminotransferase (HisC2) from Mycobacterium tuberculosis.
TLDR
HisC2 from Mycobacterium tuberculosis was overexpressed in M. smegmatis and purified to homogeneity using nickel-nitrilotriacetic acid metal-affinity and gel-filtration chromatography to record X-ray diffraction data to 2.45 Å resolution from a single crystal.Abstract:
HisC2 from Mycobacterium tuberculosis was overexpressed in M. smegmatis and purified to homogeneity using nickel–nitrilotriacetic acid metal-affinity and gel-filtration chromatography. Diffraction-quality crystals were grown using the hanging-drop vapour-diffusion technique from a condition consisting of 7 mg ml−1 HisC2 (in 20 mM Tris pH 8.8, 50 mM NaCl and 5% glycerol), 1 M succinic acid pH 7.0, 0.1 M HEPES pH 7.0 and 1%(w/v) polyethylene glycol monomethyl ether 2000. The crystals belonged to the orthorhombic space group P21212, with unit-cell parameters a = 255.98, b = 77.09, c = 117.97 A. X-ray diffraction data were recorded to 2.45 A resolution from a single crystal using the in-house X-ray facility.read more
Citations
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Journal ArticleDOI
Histidine biosynthesis, its regulation and biotechnological application in Corynebacterium glutamicum
TL;DR: This review summarizes the current knowledge of l‐histidine biosynthesis in Corynebacterium glutamicum and discusses the potential of using this knowledge to create or improve C. glutamicum strains for the industrial l‐ histidine production.
Journal ArticleDOI
Production of recombinant proteins in Mycobacterium smegmatis for structural and functional studies
Ghader Bashiri,Edward N. Baker +1 more
TL;DR: Early attempts to produce mycobacterial proteins in alternative expression hosts are described and available expression systems in M. smegmatis are focused on for developing novel and better therapeutics and diagnostics.
Journal ArticleDOI
Targeting the Histidine Pathway in Mycobacterium tuberculosis
Juleane Lunardi,José Eduardo Sacconi Nunes,Cristiano Valim Bizarro,Luiz Augusto Basso,Diógenes Santiago Santos,Pablo Machado +5 more
TL;DR: A comprehensive overview of Mycobacterium tuberculosis histidine pathway enzymes as attractive targets for the development of new antimycobacterial agents is provided, mainly summarizing the previously reported inhibition data for Mtb or orthologous proteins.
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Crystal structures of Mycobacterium tuberculosis HspAT and ArAT reveal structural basis of their distinct substrate specificities.
TL;DR: The study demonstrates that the adventitiously bound ligand 2-(N-morpholino)ethanesulfonic acid (MES) is indeed a specific inhibitor of HspAT, and suggests that previously untapped morpholine-ring scaffold compounds could be explored for the design of new anti-TB agents.
Journal ArticleDOI
Sample preparation, crystallization and structure solution of HisC from Mycobacterium tuberculosis.
TL;DR: Histidinolphosphate aminotransferase (HisC) from Mycobacterium tuberculosis was overexpressed in M. smegmatis and purified to homogeneity using nickel-nitrilotriacetic acid metal-affinity and gel-filtration chromatography.
References
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