Open AccessPurification, crystallization and preliminary crystallographic analysis of the vacuole‐type ATPase subunit E from Pyrococcus horikoshii OT3
Author(s) -
Lokanath Neratur K.,
Ukita Yoko,
Sugahara Mitsuaki,
Kunishima Naoki
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309104026430
Subject(s) - pyrococcus horikoshii , orthorhombic crystal system , protein subunit , atpase , crystallography , crystallization , escherichia coli , biology , crystal structure , chemistry , biochemistry , enzyme , organic chemistry , gene
The vacuole‐type ATPases in eukaryotic cells translocate protons across various biological membranes including the vacuolar membrane by consuming ATP molecules. The E subunit of the multisubunit complex V‐ATPase from Pyrococcus horikoshii OT3, which has a molecular weight of 22.88 kDa, has been cloned, overexpressed in Escherichia coli , purified and crystallized by the microbatch method using PEG 4000 as a precipitant at 296 K. A data set to 1.85 Å resolution with 98.8% completeness and an R merge of 6.5% was collected from a single flash‐cooled crystal using synchrotron radiation. The crystal belonged to the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 52.196, b = 55.317, c = 77.481 Å, and is most likely to contain one molecule per asymmetric unit.