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Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Enpp1.

ABSTRACT: Enpp1 is an extracellular membrane-bound glycoprotein that regulates bone mineralization by hydrolyzing ATP to generate pyrophosphate. The extracellular region of mouse Enpp1 was expressed in HEK293S GnT1(-) cells, purified using the TARGET tag/P20.1-Sepharose system and crystallized. An X-ray diffraction data set was collected to 3.0?Å resolution. The crystal belonged to space group P3(1), with unit-cell parameters a = b = 105.3, c = 173.7?Å. A single-wavelength anomalous dispersion (SAD) data set was also collected to 2.7?Å resolution using a selenomethionine-labelled crystal. The experimental phases determined by the SAD method produced an interpretable electron-density map.

SUBMITTER: Kato K

PROVIDER: S-EPMC3388920 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature

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Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Enpp1.

Kato Kazuki K Nishimasu Hiroshi H Mihara Emiko E Ishitani Ryuichiro R Takagi Junichi J Aoki Junken J Nureki Osamu O

Acta crystallographica. Section F, Structural biology and crystallization communications 20120627 Pt 7

Enpp1 is an extracellular membrane-bound glycoprotein that regulates bone mineralization by hydrolyzing ATP to generate pyrophosphate. The extracellular region of mouse Enpp1 was expressed in HEK293S GnT1(-) cells, purified using the TARGET tag/P20.1-Sepharose system and crystallized. An X-ray diffraction data set was collected to 3.0 Å resolution. The crystal belonged to space group P3(1), with unit-cell parameters a = b = 105.3, c = 173.7 Å. A single-wavelength anomalous dispersion (SAD) data ...[more]

PMID: 22750863

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