Purification, crystallization and preliminary X-ray diffraction analysis of NADP-dependent glutamate dehydrogenase from Aspergillus niger
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Title |
Purification, crystallization and preliminary X-ray diffraction analysis of NADP-dependent glutamate dehydrogenase from Aspergillus niger
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Creator |
PRAKASH, P
WALVEKAR, AS PUNEKAR, NS BHAUMIK, P |
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Subject |
CRYSTAL-STRUCTURE
ALLOSTERIC-REGULATION MECHANISM SPECIFICITY COMPLEXES FAMILY |
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Description |
Glutamate dehydrogenase (GDH) catalyzes the NAD-dependent or NADP-dependent oxidative deamination of l-glutamate to 2-oxoglutarate and ammonia. This important reversible reaction establishes the link between carbon and nitrogen metabolism. In this study, Aspergillus niger NADP-GDH (AnGDH) has been overexpressed and purified. Purified AnGDH, with a high specific activity of 631.1 units per milligram of protein, was crystallized and the crystal diffracted to 2.9 angstrom resolution using a home X-ray source. Preliminary analysis of the X-ray diffraction data showed that the crystal belonged to space group R32, with unit-cell parameters a = b = 173.8, c = 241.5 angstrom, alpha = beta = 90, gamma = 120 degrees. The crystals exhibited an unusually high solvent content (83.0%) and had only one molecule in the asymmetric unit. Initial phases were obtained by molecular replacement, and model building and structure refinement of AnGDH are in progress.
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Publisher |
WILEY-BLACKWELL
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Date |
2014-12-29T06:19:42Z
2014-12-29T06:19:42Z 2014 |
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Type |
Article
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Identifier |
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 701508-1512
1744-3091 http://dx.doi.org/10.1107/S2053230X14021499 http://dspace.library.iitb.ac.in/jspui/handle/100/17294 |
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Language |
English
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