Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1995 Apr 11;92(8):3391–3395. doi: 10.1073/pnas.92.8.3391

Transmembrane signaling characterized in bacterial chemoreceptors by using sulfhydryl cross-linking in vivo.

G F Lee 1, M R Lebert 1, A A Lilly 1, G L Hazelbauer 1
PMCID: PMC42172  PMID: 7724572

Abstract

Transmembrane signaling by bacterial chemoreceptors is thought to involve conformational changes within a stable homodimer. We investigated the functional consequences of constraining movement between pairs of helices in the four-helix structure of the transmembrane domain of chemoreceptor Trg. Using a family of cysteine-containing receptors, we identified oxidation treatments for intact cells that catalyzed essentially complete sulfhydryl cross-linking at selected positions and yet left flagellar and sensory functions largely unperturbed. Constraining movement by cross-links between subunits had little effect on tactic response, but constraining movement between transmembrane segments of the monomer drastically reduced function. We deduce that transmembrane signaling requires substantial movement between transmembrane helices of a monomer but not between interacting helices across the interface between subunits.

Full text

PDF
3391

Images in this article

3392Fig. 2
on p.3392
3394Fig. 5
on p.3394

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alex L. A., Simon M. I. Protein histidine kinases and signal transduction in prokaryotes and eukaryotes. Trends Genet. 1994 Apr;10(4):133–138. doi: 10.1016/0168-9525(94)90215-1. [DOI] [PubMed] [Google Scholar]
  2. Ames P., Chen J., Wolff C., Parkinson J. S. Structure-function studies of bacterial chemosensors. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 1):59–65. doi: 10.1101/sqb.1988.053.01.010. [DOI] [PubMed] [Google Scholar]
  3. Berg H. C., Tedesco P. M. Transient response to chemotactic stimuli in Escherichia coli. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3235–3239. doi: 10.1073/pnas.72.8.3235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bourret R. B., Borkovich K. A., Simon M. I. Signal transduction pathways involving protein phosphorylation in prokaryotes. Annu Rev Biochem. 1991;60:401–441. doi: 10.1146/annurev.bi.60.070191.002153. [DOI] [PubMed] [Google Scholar]
  5. Burrows G. G., Newcomer M. E., Hazelbauer G. L. Purification of receptor protein Trg by exploiting a property common to chemotactic transducers of Escherichia coli. J Biol Chem. 1989 Oct 15;264(29):17309–17315. [PubMed] [Google Scholar]
  6. Danielson M. A., Biemann H. P., Koshland D. E., Jr, Falke J. J. Attractant- and disulfide-induced conformational changes in the ligand binding domain of the chemotaxis aspartate receptor: a 19F NMR study. Biochemistry. 1994 May 24;33(20):6100–6109. doi: 10.1021/bi00186a009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Falke J. J., Dernburg A. F., Sternberg D. A., Zalkin N., Milligan D. L., Koshland D. E., Jr Structure of a bacterial sensory receptor. A site-directed sulfhydryl study. J Biol Chem. 1988 Oct 15;263(29):14850–14858. [PubMed] [Google Scholar]
  8. Falke J. J., Koshland D. E., Jr Global flexibility in a sensory receptor: a site-directed cross-linking approach. Science. 1987 Sep 25;237(4822):1596–1600. doi: 10.1126/science.2820061. [DOI] [PubMed] [Google Scholar]
  9. Francis G., Brennan L., Ferenci T. Affinity-chromatographic purification of sixteen cysteine-substituted maltoporin variants: thiol reactivity and cross-linking in an outer membrane protein of Escherichia coli. Biochim Biophys Acta. 1991 Aug 5;1067(1):89–96. doi: 10.1016/0005-2736(91)90029-8. [DOI] [PubMed] [Google Scholar]
  10. Gegner J. A., Graham D. R., Roth A. F., Dahlquist F. W. Assembly of an MCP receptor, CheW, and kinase CheA complex in the bacterial chemotaxis signal transduction pathway. Cell. 1992 Sep 18;70(6):975–982. doi: 10.1016/0092-8674(92)90247-a. [DOI] [PubMed] [Google Scholar]
  11. Hazelbauer G. L., Harayama S. Mutants in transmission of chemotactic signals from two independent receptors of E. coli. Cell. 1979 Mar;16(3):617–625. doi: 10.1016/0092-8674(79)90035-7. [DOI] [PubMed] [Google Scholar]
  12. Lee G. F., Burrows G. G., Lebert M. R., Dutton D. P., Hazelbauer G. L. Deducing the organization of a transmembrane domain by disulfide cross-linking. The bacterial chemoreceptor Trg. J Biol Chem. 1994 Nov 25;269(47):29920–29927. [PubMed] [Google Scholar]
  13. Lynch B. A., Koshland D. E., Jr Disulfide cross-linking studies of the transmembrane regions of the aspartate sensory receptor of Escherichia coli. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10402–10406. doi: 10.1073/pnas.88.23.10402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Milburn M. V., Privé G. G., Milligan D. L., Scott W. G., Yeh J., Jancarik J., Koshland D. E., Jr, Kim S. H. Three-dimensional structures of the ligand-binding domain of the bacterial aspartate receptor with and without a ligand. Science. 1991 Nov 29;254(5036):1342–1347. doi: 10.1126/science.1660187. [DOI] [PubMed] [Google Scholar]
  15. Milligan D. L., Koshland D. E., Jr Intrasubunit signal transduction by the aspartate chemoreceptor. Science. 1991 Dec 13;254(5038):1651–1654. doi: 10.1126/science.1661030. [DOI] [PubMed] [Google Scholar]
  16. Milligan D. L., Koshland D. E., Jr Site-directed cross-linking. Establishing the dimeric structure of the aspartate receptor of bacterial chemotaxis. J Biol Chem. 1988 May 5;263(13):6268–6275. [PubMed] [Google Scholar]
  17. Morgan D. G., Baumgartner J. W., Hazelbauer G. L. Proteins antigenically related to methyl-accepting chemotaxis proteins of Escherichia coli detected in a wide range of bacterial species. J Bacteriol. 1993 Jan;175(1):133–140. doi: 10.1128/jb.175.1.133-140.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pakula A. A., Simon M. I. Determination of transmembrane protein structure by disulfide cross-linking: the Escherichia coli Tar receptor. Proc Natl Acad Sci U S A. 1992 May 1;89(9):4144–4148. doi: 10.1073/pnas.89.9.4144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Park C., Hazelbauer G. L. Mutations specifically affecting ligand interaction of the Trg chemosensory transducer. J Bacteriol. 1986 Jul;167(1):101–109. doi: 10.1128/jb.167.1.101-109.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Parkinson J. S. Signal transduction schemes of bacteria. Cell. 1993 Jun 4;73(5):857–871. doi: 10.1016/0092-8674(93)90267-t. [DOI] [PubMed] [Google Scholar]
  21. Schuster S. C., Swanson R. V., Alex L. A., Bourret R. B., Simon M. I. Assembly and function of a quaternary signal transduction complex monitored by surface plasmon resonance. Nature. 1993 Sep 23;365(6444):343–347. doi: 10.1038/365343a0. [DOI] [PubMed] [Google Scholar]
  22. Srinivasan N., Sowdhamini R., Ramakrishnan C., Balaram P. Conformations of disulfide bridges in proteins. Int J Pept Protein Res. 1990 Aug;36(2):147–155. doi: 10.1111/j.1399-3011.1990.tb00958.x. [DOI] [PubMed] [Google Scholar]
  23. Stoddard B. L., Bui J. D., Koshland D. E., Jr Structure and dynamics of transmembrane signaling by the Escherichia coli aspartate receptor. Biochemistry. 1992 Dec 8;31(48):11978–11983. doi: 10.1021/bi00163a004. [DOI] [PubMed] [Google Scholar]
  24. Yaghmai R., Hazelbauer G. L. Ligand occupancy mimicked by single residue substitutions in a receptor: transmembrane signaling induced by mutation. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7890–7894. doi: 10.1073/pnas.89.17.7890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yaghmai R., Hazelbauer G. L. Strategies for differential sensory responses mediated through the same transmembrane receptor. EMBO J. 1993 May;12(5):1897–1905. doi: 10.1002/j.1460-2075.1993.tb05838.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yeh J. I., Biemann H. P., Pandit J., Koshland D. E., Kim S. H. The three-dimensional structure of the ligand-binding domain of a wild-type bacterial chemotaxis receptor. Structural comparison to the cross-linked mutant forms and conformational changes upon ligand binding. J Biol Chem. 1993 May 5;268(13):9787–9792. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES