Open AccessExpression, purification, crystallization and preliminary X‐ray crystallographic analysis of Enpp1
Author(s) -
Kato Kazuki,
Nishimasu Hiroshi,
Mihara Emiko,
Ishitani Ryuichiro,
Takagi Junichi,
Aoki Junken,
Nureki Osamu
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112019306
Subject(s) - crystallization , extracellular , crystallography , resolution (logic) , pyrophosphate , chemistry , crystal (programming language) , membrane , x ray , crystal structure , materials science , physics , enzyme , optics , biochemistry , organic chemistry , artificial intelligence , computer science , programming language
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 P 3 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.