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

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

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

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 P31, 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.

Original languageEnglish
Pages (from-to)778-782
Number of pages5
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume68
Issue number7
DOIs
Publication statusPublished - 2012 Jul

Keywords

  • Enpp1

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

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