Use of Random and Site-Directed Mutagenesis to Probe Protein Structure-Function Relationships: Applied Techniques in the Study of Helicobacter pylori

Jeannette M Whitmire; D Scott Merrell

doi:10.1007/978-1-4939-6472-7_31

Use of Random and Site-Directed Mutagenesis to Probe Protein Structure-Function Relationships: Applied Techniques in the Study of Helicobacter pylori

Methods Mol Biol. 2017:1498:461-480. doi: 10.1007/978-1-4939-6472-7_31.

Authors

Jeannette M Whitmire¹, D Scott Merrell²

Affiliations

¹ Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA.
² Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD, 20814, USA. douglas.merrell@usuhs.edu.

PMID: 27709595
DOI: 10.1007/978-1-4939-6472-7_31

Abstract

Mutagenesis is a valuable tool to examine the structure-function relationships of bacterial proteins. As such, a wide variety of mutagenesis techniques and strategies have been developed. This chapter details a selection of random mutagenesis methods and site-directed mutagenesis procedures that can be applied to an array of bacterial species. Additionally, the direct application of the techniques to study the Helicobacter pylori Ferric Uptake Regulator (Fur) protein is described. The varied approaches illustrated herein allow the robust investigation of the structural-functional relationships within a protein of interest.

Keywords: Bacterial mutagenesis; Protein analysis; Random mutagenesis; Site-directed mutagenesis; Structure–function.

MeSH terms

Bacterial Proteins / genetics*
Helicobacter pylori / genetics*
Iron / metabolism
Mutagenesis / genetics*
Mutagenesis, Site-Directed / methods
Repressor Proteins / genetics
Structure-Activity Relationship

Substances

Bacterial Proteins
Repressor Proteins
Iron