Article

Crystallization and preliminary X-ray characterization of phosphoglucose isomerase from Mycobacterium tuberculosis H37Rv.

Gene Regulation Laboratory, National Institute of Immunology, New Delhi 110067, India.
Acta Crystallographica Section F Structural Biology and Crystallization Communications (impact factor: 0.51). 04/2007; 63(Pt 4):353-5. DOI:10.1107/S1744309107013218 pp.353-5
Source: PubMed

ABSTRACT Phosphoglucose isomerase is a ubiquitous enzyme that catalyzes the isomerization of D-glucopyranose-6-phosphate to D-fructofuranose-6-phosphate. The present investigation reports the expression, purification, crystallization and preliminary crystallographic studies of the phosphoglucose isomerase from Mycobacterium tuberculosis H37Rv, which shares 46% sequence identity with that of its human host. The recombinant protein, which was prepared using an Escherichia coli expression system, was crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to a resolution of 2.8 A and belonged to the orthorhombic space group I2(1)2(1)2(1), with unit-cell parameters a = 109.0, b = 119.8, c = 138.9 A.

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  • Article: A phosphoglucose isomerase gene is involved in the Rag phenotype of the yeast Kluyveromyces lactis.
    P Goffrini, M Wésolowski-Louvel, I Ferrero
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    ABSTRACT: The rag2 mutant of Kluyveromyces lactis cannot grow on glucose when mitochondrial functions are blocked by various mitochondrial inhibitors, suggesting the presence of a defect in the fermentation pathway. The RAG2 gene has been cloned from a K. lactis genomic library by complementation of the rag2 mutation. The amino acid sequence of the RAG2 protein deduced from the nucleotide sequence of the cloned RAG2 gene shows homology to the sequences of known phosphoglucose isomerases (PGI and PHI). In vivo complementation of the pgi1 mutation in Saccharomyces cerevisiae by the cloned RAG2 gene, together with measurements of specific PGI activities and the detection of PGI proteins, confirm that the RAG2 gene of K. lactis codes for the phosphoglucose isomerase enzyme. Complete loss of PGI activity observed when the coding sequence of RAG2 was disrupted leads us to conclude that RAG2 is the only gene that codes for phosphoglucose isomerase in K. lactis. The RAG2 gene of K. lactis is expressed constitutively, independently of the growth substrates (glycolytic or gluconeogenic). Unlike the pgi1 mutants of S. cerevisiae, the K. lactis rag2 mutants can still grow on glucose, however they do not produce ethanol.
    MGG - Molecular and General Genetics 10/1991; 228(3):401-9.
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Keywords

Escherichia coli expression system
 
hanging-drop vapour-diffusion method
 
Mycobacterium tuberculosis H37Rv
 
preliminary crystallographic studies
 
present investigation reports
 
recombinant protein
 
shares 46% sequence identity
 
ubiquitous enzyme