Open AccessCloning, purification and preliminary crystallographic analysis of the complex of Helicobacter pylori α‐carbonic anhydrase with acetazolamide
Author(s) -
Modak Joyanta K.,
RevittMills Sarah A.,
Roujeinikova Anna
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113026146
Subject(s) - periplasmic space , helicobacter pylori , acetazolamide , carbonic anhydrase , chemistry , urease , biochemistry , crystallography , biology , enzyme , escherichia coli , medicine , gene , genetics
Helicobacter pylori infection of the stomach can lead to severe gastroduodenal diseases such as gastritis, peptic ulcers and gastric cancers. Periplasmic H. pylori α‐carbonic anhydrase (HpαCA) is essential for the acclimatization of the bacterium to the acidity of the stomach. Through the action of urease and carbonic anhydrases, the H. pylori periplasmic pH is maintained at around 6 in an environment with a pH as low as 2, which in turn facilitates the maintenance of a cytoplasmic pH close to neutral, allowing growth in the gastric niche. Crystals of HpαCA in complex with the inhibitor acetazolamide have been grown by the hanging‐drop vapour‐diffusion method using polyethylene glycol as a precipitating agent. The crystals have the symmetry of space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 37.0, b = 82.4, c = 150.8 Å. An X‐ray diffraction data set was collected from a single crystal to 1.7 Å resolution. Calculation of the self‐rotation function using this data and molecular replacement showed that the asymmetric unit contains an HpαCA dimer.