The purification, crystallization and preliminary diffraction of a glycerophosphodiesterase from Enterobacter aerogenes

Colin J. Jackson; Paul D. Carr; Hye Kyung Kim; Jian Wei Liu; David L. Ollis

doi:10.1107/S1744309106020021

The purification, crystallization and preliminary diffraction of a glycerophosphodiesterase from Enterobacter aerogenes

Colin J. Jackson, Paul D. Carr, Hye Kyung Kim, Jian Wei Liu, David L. Ollis^*

^*Corresponding author for this work

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8 Citations (Scopus)

Abstract

The metallo-glycerophosphodiesterase from Enterobacter aerogenes (GpdQ) has been cloned, expressed in Escherichia coli and purified. Initial screening of crystallization conditions for this enzyme resulted in the identification of needles from one condition in a sodium malonate grid screen. Removal of the metals from the enzyme and subsequent optimization of these conditions led to crystals that diffracted to 2.9 Å and belonged to space group P2 ₁3, with unit-cell parameter a = 164.1 Å. Self-rotation function analysis and V_M calculations indicated that the asymmetric unit contains two copies of the monomeric enzyme, corresponding to a solvent content of 79%. It is intended to determine the structure of this protein utilizing SAD phasing from transition metals or molecular replacement.

Original language	English
Pages (from-to)	659-661
Number of pages	3
Journal	Acta Crystallographica Section F: Structural Biology and Crystallization Communications
Volume	62
Issue number	7
DOIs	https://doi.org/10.1107/S1744309106020021
Publication status	Published - Jul 2006

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title = "The purification, crystallization and preliminary diffraction of a glycerophosphodiesterase from Enterobacter aerogenes",

abstract = "The metallo-glycerophosphodiesterase from Enterobacter aerogenes (GpdQ) has been cloned, expressed in Escherichia coli and purified. Initial screening of crystallization conditions for this enzyme resulted in the identification of needles from one condition in a sodium malonate grid screen. Removal of the metals from the enzyme and subsequent optimization of these conditions led to crystals that diffracted to 2.9 {\AA} and belonged to space group P2 13, with unit-cell parameter a = 164.1 {\AA}. Self-rotation function analysis and VM calculations indicated that the asymmetric unit contains two copies of the monomeric enzyme, corresponding to a solvent content of 79%. It is intended to determine the structure of this protein utilizing SAD phasing from transition metals or molecular replacement.",

author = "Jackson, {Colin J.} and Carr, {Paul D.} and Kim, {Hye Kyung} and Liu, {Jian Wei} and Ollis, {David L.}",

year = "2006",

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journal = "Acta Crystallographica Section F: Structural Biology and Crystallization Communications",

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The purification, crystallization and preliminary diffraction of a glycerophosphodiesterase from Enterobacter aerogenes. / Jackson, Colin J.; Carr, Paul D.; Kim, Hye Kyung et al.
In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 62, No. 7, 07.2006, p. 659-661.

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

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AU - Ollis, David L.

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AB - The metallo-glycerophosphodiesterase from Enterobacter aerogenes (GpdQ) has been cloned, expressed in Escherichia coli and purified. Initial screening of crystallization conditions for this enzyme resulted in the identification of needles from one condition in a sodium malonate grid screen. Removal of the metals from the enzyme and subsequent optimization of these conditions led to crystals that diffracted to 2.9 Å and belonged to space group P2 13, with unit-cell parameter a = 164.1 Å. Self-rotation function analysis and VM calculations indicated that the asymmetric unit contains two copies of the monomeric enzyme, corresponding to a solvent content of 79%. It is intended to determine the structure of this protein utilizing SAD phasing from transition metals or molecular replacement.

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

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The purification, crystallization and preliminary diffraction of a glycerophosphodiesterase from Enterobacter aerogenes

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