Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Crystallization and X‐ray analysis of the extracellular adhesion domain of Helicobacter pylori adhesin A: the significance of the cation composition in the crystallization precipitant

Crystallization and X‐ray analysis of the extracellular adhesion domain of Helicobacter pylori... Adherence to host cells is a crucial step in the process of bacterial infection, which is usually mediated by a number of outer membrane proteins identified as adhesins. Helicobacter pylori adhesin A (HpaA) is a member of the adhesin family that mediates the adherence of Helicobacter pylori to gastric epithelial cells, and consequently assists the bacteria in becoming a life‐long colonizer of the human stomach. In this study, two constructs were made for the production of truncated HpaA proteins comprising residues 31–260 and 53–260, respectively. The products of both constructs were crystallized, but only the protein from the shorter construct (residues 53–260) formed crystals that were capable of diffraction. In the subsequent optimization trials, crystals in different forms were unexpectedly obtained by using lithium sulfate and ammonium sulfate as the precipitant. An X‐ray data set was collected to 1.95 Å resolution on beamline BL18U1 at SSRF using a crystal grown with 1.92 M lithium sulfate, which belonged to space group P65 with unit‐cell parameters a = b = 95.42, c = 54.72 Å, γ = 120°, while another crystal grown with 1.9 M ammonium sulfate diffracted to 2.60 Å resolution and the collected data set was indexed in space group P21212, with unit‐cell parameters a = 121.01, b = 190.56, c = 106.31 Å. The collection of diffraction data has established a solid basis for structure determination. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Acta Crystallographica Section F Wiley

Crystallization and X‐ray analysis of the extracellular adhesion domain of Helicobacter pylori adhesin A: the significance of the cation composition in the crystallization precipitant

Download PDF
7 pages

Loading next page...
 
/lp/wiley/crystallization-and-x-ray-analysis-of-the-extracellular-adhesion-rlmxFTw0ZW

References (26)

Publisher
Wiley
Copyright
Copyright © 2017 Wiley Subscription Services
ISSN
2053-230X
eISSN
2053-230X
DOI
10.1107/S2053230X17003004
pmid
28368278
Publisher site
See Article on Publisher Site

Abstract

Adherence to host cells is a crucial step in the process of bacterial infection, which is usually mediated by a number of outer membrane proteins identified as adhesins. Helicobacter pylori adhesin A (HpaA) is a member of the adhesin family that mediates the adherence of Helicobacter pylori to gastric epithelial cells, and consequently assists the bacteria in becoming a life‐long colonizer of the human stomach. In this study, two constructs were made for the production of truncated HpaA proteins comprising residues 31–260 and 53–260, respectively. The products of both constructs were crystallized, but only the protein from the shorter construct (residues 53–260) formed crystals that were capable of diffraction. In the subsequent optimization trials, crystals in different forms were unexpectedly obtained by using lithium sulfate and ammonium sulfate as the precipitant. An X‐ray data set was collected to 1.95 Å resolution on beamline BL18U1 at SSRF using a crystal grown with 1.92 M lithium sulfate, which belonged to space group P65 with unit‐cell parameters a = b = 95.42, c = 54.72 Å, γ = 120°, while another crystal grown with 1.9 M ammonium sulfate diffracted to 2.60 Å resolution and the collected data set was indexed in space group P21212, with unit‐cell parameters a = 121.01, b = 190.56, c = 106.31 Å. The collection of diffraction data has established a solid basis for structure determination.

Journal

Acta Crystallographica Section F – Wiley

Published: Jan 1, 2017

Keywords: ; ; ; ;

There are no references for this article.