Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Enpp6

Junko Morita; Kazuki Kato; Emiko Mihara; Ryuichiro Ishitani; Junichi Takagi; Hiroshi Nishimasu; Junken Aoki; Osamu Nureki

doi:10.1107/S2053230X14008929

Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Enpp6

Junko Morita, Kazuki Kato, Emiko Mihara, Ryuichiro Ishitani, Junichi Takagi, Hiroshi Nishimasu, Junken Aoki, Osamu Nureki

PHA - Life and Pharmaceutical Science

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

1 Citation (Scopus)

Abstract

Enpp (ectonucleotide phosphodiesterase/pyrophosphatase) 6 is a membrane-bound glycoprotein that hydrolyzes choline-containing compounds such as lysophosphatidylcholine and glycerophosphorylcholine, and presumably participates in choline metabolism. The catalytic domain of mouse Enpp6 was expressed in HEK293T cells, purified using the TARGET tag/P20.1-Sepharose system and crystallized. An X-ray diffraction data set was collected to 1.8 Å resolution. The crystal belonged to space group P1, with unit-cell parameters a = 63.7, b = 68.8, c = 69.7 Å, α = 60.6, β = 87.0, γ = 68.1°. Assuming the presence of two protein molecules per asymmetric unit, the solvent content was estimated to be 49.5%.

Original language	English
Pages (from-to)	794-799
Number of pages	6
Journal	Acta Crystallographica Section F:Structural Biology Communications
Volume	70
Issue number	6
DOIs	https://doi.org/10.1107/S2053230X14008929
Publication status	Published - 2014 Jun

Keywords

Enpp6
N-glycosylation
choline

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Genetics
Condensed Matter Physics

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Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Enpp6. / Morita, Junko; Kato, Kazuki; Mihara, Emiko; Ishitani, Ryuichiro; Takagi, Junichi; Nishimasu, Hiroshi; Aoki, Junken; Nureki, Osamu.

In: Acta Crystallographica Section F:Structural Biology Communications, Vol. 70, No. 6, 06.2014, p. 794-799.

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

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abstract = "Enpp (ectonucleotide phosphodiesterase/pyrophosphatase) 6 is a membrane-bound glycoprotein that hydrolyzes choline-containing compounds such as lysophosphatidylcholine and glycerophosphorylcholine, and presumably participates in choline metabolism. The catalytic domain of mouse Enpp6 was expressed in HEK293T cells, purified using the TARGET tag/P20.1-Sepharose system and crystallized. An X-ray diffraction data set was collected to 1.8 {\AA} resolution. The crystal belonged to space group P1, with unit-cell parameters a = 63.7, b = 68.8, c = 69.7 {\AA}, α = 60.6, β = 87.0, γ = 68.1°. Assuming the presence of two protein molecules per asymmetric unit, the solvent content was estimated to be 49.5%.",

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AU - Takagi, Junichi

AU - Nishimasu, Hiroshi

AU - Aoki, Junken

AU - Nureki, Osamu

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AB - Enpp (ectonucleotide phosphodiesterase/pyrophosphatase) 6 is a membrane-bound glycoprotein that hydrolyzes choline-containing compounds such as lysophosphatidylcholine and glycerophosphorylcholine, and presumably participates in choline metabolism. The catalytic domain of mouse Enpp6 was expressed in HEK293T cells, purified using the TARGET tag/P20.1-Sepharose system and crystallized. An X-ray diffraction data set was collected to 1.8 Å resolution. The crystal belonged to space group P1, with unit-cell parameters a = 63.7, b = 68.8, c = 69.7 Å, α = 60.6, β = 87.0, γ = 68.1°. Assuming the presence of two protein molecules per asymmetric unit, the solvent content was estimated to be 49.5%.

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