Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5′-phosphate oxidase from Mycobacterium smegmatis

Colin J. Jackson; Matthew C. Taylor; David B. Tattersall; Nigel G. French; Paul D. Carr; David L. Ollis; Robyn J. Russell; John G. Oakeshott

doi:10.1107/S1744309108011512

Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5′-phosphate oxidase from Mycobacterium smegmatis

Colin J. Jackson, Matthew C. Taylor, David B. Tattersall, Nigel G. French, Paul D. Carr, David L. Ollis, Robyn J. Russell, John G. Oakeshott

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

Abstract

Pyridoxine 5′-phosphate oxidases (PNPOxs) are known to catalyse the terminal step in pyridoxal 5′-phosphate biosynthesis in a flavin mononucleotide-dependent manner in humans and Escherichia coli. Recent reports of a putative PNPOx from Mycobacterium tuberculosis, Rv1155, suggest that the cofactor or catalytic mechanism may differ in Mycobacterium species. To investigate this, a putative PNPOx from M. smegmatis, Msmeg_3380, has been cloned. This enzyme has been recombinantly expressed in E. coli and purified to homogeneity. Good-quality crystals of selenomethionine-substituted Msmeg_3380 were obtained by the hanging-drop vapour-diffusion technique and diffracted to 1.2 Å using synchrotron radiation.

Original language	English
Pages (from-to)	435-437
Number of pages	3
Journal	Acta Crystallographica Section F: Structural Biology and Crystallization Communications
Volume	64
Issue number	5
DOIs	https://doi.org/10.1107/S1744309108011512
Publication status	Published - 5 Apr 2008

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Jackson, C. J., Taylor, M. C., Tattersall, D. B., French, N. G., Carr, P. D., Ollis, D. L., Russell, R. J., & Oakeshott, J. G. (2008). Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5′-phosphate oxidase from Mycobacterium smegmatis. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 64(5), 435-437. https://doi.org/10.1107/S1744309108011512

@article{ef45c2cf08754c6581f6b32ed9a41888,

title = "Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5′-phosphate oxidase from Mycobacterium smegmatis",

abstract = "Pyridoxine 5′-phosphate oxidases (PNPOxs) are known to catalyse the terminal step in pyridoxal 5′-phosphate biosynthesis in a flavin mononucleotide-dependent manner in humans and Escherichia coli. Recent reports of a putative PNPOx from Mycobacterium tuberculosis, Rv1155, suggest that the cofactor or catalytic mechanism may differ in Mycobacterium species. To investigate this, a putative PNPOx from M. smegmatis, Msmeg_3380, has been cloned. This enzyme has been recombinantly expressed in E. coli and purified to homogeneity. Good-quality crystals of selenomethionine-substituted Msmeg_3380 were obtained by the hanging-drop vapour-diffusion technique and diffracted to 1.2 {\AA} using synchrotron radiation.",

keywords = "FMN, Mycobacterium smegmatis, Pyridoxine 5′-phosphate oxidases",

author = "Jackson, {Colin J.} and Taylor, {Matthew C.} and Tattersall, {David B.} and French, {Nigel G.} and Carr, {Paul D.} and Ollis, {David L.} and Russell, {Robyn J.} and Oakeshott, {John G.}",

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Jackson, CJ, Taylor, MC, Tattersall, DB, French, NG, Carr, PD, Ollis, DL, Russell, RJ & Oakeshott, JG 2008, 'Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5′-phosphate oxidase from Mycobacterium smegmatis', Acta Crystallographica Section F: Structural Biology and Crystallization Communications, vol. 64, no. 5, pp. 435-437. https://doi.org/10.1107/S1744309108011512

Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5′-phosphate oxidase from Mycobacterium smegmatis. / Jackson, Colin J.; Taylor, Matthew C.; Tattersall, David B. et al.
In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 64, No. 5, 05.04.2008, p. 435-437.

Research output: Contribution to journal › Article › peer-review

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T1 - Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5′-phosphate oxidase from Mycobacterium smegmatis

AU - Jackson, Colin J.

AU - Taylor, Matthew C.

AU - Tattersall, David B.

AU - French, Nigel G.

AU - Carr, Paul D.

AU - Ollis, David L.

AU - Russell, Robyn J.

AU - Oakeshott, John G.

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AB - Pyridoxine 5′-phosphate oxidases (PNPOxs) are known to catalyse the terminal step in pyridoxal 5′-phosphate biosynthesis in a flavin mononucleotide-dependent manner in humans and Escherichia coli. Recent reports of a putative PNPOx from Mycobacterium tuberculosis, Rv1155, suggest that the cofactor or catalytic mechanism may differ in Mycobacterium species. To investigate this, a putative PNPOx from M. smegmatis, Msmeg_3380, has been cloned. This enzyme has been recombinantly expressed in E. coli and purified to homogeneity. Good-quality crystals of selenomethionine-substituted Msmeg_3380 were obtained by the hanging-drop vapour-diffusion technique and diffracted to 1.2 Å using synchrotron radiation.

KW - FMN

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JO - Acta Crystallographica Section F: Structural Biology and Crystallization Communications

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Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5′-phosphate oxidase from Mycobacterium smegmatis

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