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Improved expression and purification of the Helicobacter pylori adhesin BabA through the incorporation of a hexa-lysine tag

Hage, Naim; Falcone, Franco; Renshaw, Jonathan G.; Sebastiaan Winkler, G.; Gellert, Paul; Stolnik, Snow

Authors

Naim Hage

Franco Falcone

Jonathan G. Renshaw

Paul Gellert

Snow Stolnik



Abstract

Helicobacter pylori is a pathogenic bacterium that has the remarkable ability to withstand the harsh conditions of the stomach for decades. This is achieved through unique evolutionary adaptations, which include binding Lewisb antigens found on the gastric epithelium using the outer membrane protein BabA. We show here the yield of a recombinant form of BabA, comprising its putative extracellular binding domain, can be significantly increased through the addition of a hexa-lysine tag to the C-terminus of the protein. BabA was expressed in the periplasmic space of Escherichiacoli and purified using immobilised metal ion affinity and size exclusion chromatography – yielding approximately 1.8 mg of protein per litre of culture. The hexa-lysine tag does not inhibit the binding activity of BabA as the recombinant protein was found to possess affinity towards HSA–Lewisb glycoconjugates.

Citation

Hage, N., Falcone, F., Renshaw, J. G., Sebastiaan Winkler, G., Gellert, P., & Stolnik, S. (2015). Improved expression and purification of the Helicobacter pylori adhesin BabA through the incorporation of a hexa-lysine tag. Protein Expression and Purification, 106, 25-30. doi:10.1016/j.pep.2014.10.009

Journal Article Type Article
Acceptance Date Oct 25, 2014
Online Publication Date Oct 25, 2014
Publication Date Feb 1, 2015
Deposit Date May 24, 2018
Publicly Available Date Mar 28, 2024
Electronic ISSN 1096-0279
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 106
Pages 25-30
DOI https://doi.org/10.1016/j.pep.2014.10.009
Public URL https://nottingham-repository.worktribe.com/output/1101048
Publisher URL https://www.sciencedirect.com/science/article/pii/S1046592814002344?via%3Dihub
PMID 25448827
Additional Information eStaffProfile Description: , eStaffProfile Brief Description of Type:

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