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Cloning, purification and preliminary crystallographic analysis of the complex of Helicobacter pylori α‐carbonic anhydrase with acetazolamide

Cloning, purification and preliminary crystallographic analysis of the complex of Helicobacter... Helicobacter pylori infection of the stomach can lead to severe gastroduodenal diseases such as gastritis, peptic ulcers and gastric cancers. Periplasmic H. pyloriα‐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 P212121, 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Acta Crystallographica Section F Wiley

Cloning, purification and preliminary crystallographic analysis of the complex of Helicobacter pylori α‐carbonic anhydrase with acetazolamide

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References (41)

Publisher
Wiley
Copyright
International Union of Crystallography, 2013
ISSN
1744-3091
eISSN
1744-3091
DOI
10.1107/S1744309113026146
pmid
24192362
Publisher site
See Article on Publisher Site

Abstract

Helicobacter pylori infection of the stomach can lead to severe gastroduodenal diseases such as gastritis, peptic ulcers and gastric cancers. Periplasmic H. pyloriα‐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 P212121, 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.

Journal

Acta Crystallographica Section FWiley

Published: Nov 1, 2013

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