Abstract
Interactions between ligands and chemoreceptors in Escherichia coli and Salmonella enterica can be studied through genetic manipulation of the actors involved. Sequence analysis and modeling can reveal potential sites of interaction, and these sites can be deleted or mutated and the effects tested through various in vivo chemotaxis assays to ascertain their importance during interaction. Here, the approach for analysis of the interaction between a major E. coli chemoreceptor and its binding protein ligand is described.
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References
Krikos A, Conley MP, Boyd A, Berg HC, Simon MI (1985) Chimeric chemosensory transducers of Escherichia coli. Proc Natl Acad Sci U S A 82:1326–1330
Mowbray SL, Koshland DE Jr (1990) Mutations in the aspartate receptor of Escherichia coli which affect aspartate binding. J Biol Chem 265:15638–15643
Hazelbauer GL, Engstrom P (1980) Parallel pathways for transduction of chemotactic signals in Escherichia coli. Nature 283:98–100
Englert DL, Adase CA, Jayaraman A, Manson MD (2010) Repellent taxis in response to nickel ion requires neither Ni2+ transport nor the periplasmic NikA binding protein. J Bacteriol 192:2633–2637
Bowie JU, Pakula AA, Simon MI (1995) The three-dimensional structure of the aspartate receptor from Escherichia Coli. Acta Crystallogr Sect D Biol Crystallogr 51:145–154
Milburn MV, Prive GG, Milligan DL, Scott WG, Yeh J et al (1991) Three-dimensional structures of the ligand-binding domain of the bacterial aspartate receptor with and without a ligand. Science 254:1342–1347
Scott WG, Milligan DL, Milburn MV, Prive GG, Yeh J et al (1993) Refined structures of the ligand-binding domain of the aspartate receptor from salmonella typhimurium. J Mol Biol 232:555–573
Gardina P, Conway C, Kossman M, Manson M (1992) Aspartate and maltose-binding protein interact with adjacent sites in the tar chemotactic signal transducer of Escherichia coli. J Bacteriol 174:1528–1536
Zhang Y, Gardina PJ, Kuebler AS, Kang HS, Christopher JA et al (1999) Model of maltose-binding protein/chemoreceptor complex supports intrasubunit signaling mechanism. Proc Natl Acad Sci U S A 96:939–944
Carter P (1986) Site-directed mutagenesis. Biochem J 237:1–7
Hegde M, Englert DL, Schrock S, Cohn WB, Vogt C et al (2011) Chemotaxis to the quorum-sensing signal AI-2 requires the Tsr chemoreceptor and the periplasmic LsrB AI-2-binding protein. J Bacteriol 193:768–773
Tajima H, Imada K, Sakuma M, Hattori F, Nara T et al (2011) Ligand specificity determined by differentially arranged common ligand-binding residues in bacterial amino acid chemoreceptors Tsr and Tar. J Biol Chem 286:42200–42210
Miller ST, Xavier KB, Campagna SR, Taga ME, Semmelhack MF et al (2004) Salmonella typhimurium recognizes a chemically distinct form of the bacterial quorum-sensing signal AI-2. Mol Cell 15:677–687
Acknowledgment
I would like to acknowledge the guidance of Dr. Michael Manson over the course of my research on bacterial chemotaxis. I would also like to acknowledge the student and postdoctoral members of the Manson Lab at Texas A&M University for their hard work and collaboration with these research endeavors.
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Seely, A.L. (2018). Mutational Analysis of Binding Protein–Chemoreceptor Interactions. In: Manson, M. (eds) Bacterial Chemosensing. Methods in Molecular Biology, vol 1729. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7577-8_9
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DOI: https://doi.org/10.1007/978-1-4939-7577-8_9
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