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. 1996 Jan;178(1):289–292. doi: 10.1128/jb.178.1.289-292.1996

Purification and in vitro phosphorylation of Myxococcus xanthus AsgA protein.

Y Li 1, L Plamann 1
PMCID: PMC177652  PMID: 8550431

Abstract

The deduced amino acid sequence of the Myxococcus xanthus AsgA protein contains an N-terminal domain that is homologous to the receiver of response regulators and a C-terminal domain that is homologous to the transmitter of histidine protein kinases. We overexpressed affinity-tagged AsgA in Escherichia coli, purified the recombinant protein, and showed that AsgA has autokinase activity in vitro. The results of chemical-stability assays suggest that AsgA is phosphorylated on a histidine and provide no evidence for transfer of the phosphoryl group to the conserved aspartate of the receiver domain.

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Selected References

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  1. Downard J., Ramaswamy S. V., Kil K. S. Identification of esg, a genetic locus involved in cell-cell signaling during Myxococcus xanthus development. J Bacteriol. 1993 Dec;175(24):7762–7770. doi: 10.1128/jb.175.24.7762-7770.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Duclos B., Marcandier S., Cozzone A. J. Chemical properties and separation of phosphoamino acids by thin-layer chromatography and/or electrophoresis. Methods Enzymol. 1991;201:10–21. doi: 10.1016/0076-6879(91)01004-l. [DOI] [PubMed] [Google Scholar]
  3. Hagen D. C., Bretscher A. P., Kaiser D. Synergism between morphogenetic mutants of Myxococcus xanthus. Dev Biol. 1978 Jun;64(2):284–296. doi: 10.1016/0012-1606(78)90079-9. [DOI] [PubMed] [Google Scholar]
  4. Iuchi S., Lin E. C. Purification and phosphorylation of the Arc regulatory components of Escherichia coli. J Bacteriol. 1992 Sep;174(17):5617–5623. doi: 10.1128/jb.174.17.5617-5623.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kroos L., Kaiser D. Expression of many developmentally regulated genes in Myxococcus depends on a sequence of cell interactions. Genes Dev. 1987 Oct;1(8):840–854. doi: 10.1101/gad.1.8.840. [DOI] [PubMed] [Google Scholar]
  6. Kroos L., Kuspa A., Kaiser D. A global analysis of developmentally regulated genes in Myxococcus xanthus. Dev Biol. 1986 Sep;117(1):252–266. doi: 10.1016/0012-1606(86)90368-4. [DOI] [PubMed] [Google Scholar]
  7. Kuspa A., Plamann L., Kaiser D. A-signalling and the cell density requirement for Myxococcus xanthus development. J Bacteriol. 1992 Nov;174(22):7360–7369. doi: 10.1128/jb.174.22.7360-7369.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kuspa A., Plamann L., Kaiser D. Identification of heat-stable A-factor from Myxococcus xanthus. J Bacteriol. 1992 May;174(10):3319–3326. doi: 10.1128/jb.174.10.3319-3326.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. McCleary W. R., Zusman D. R. Purification and characterization of the Myxococcus xanthus FrzE protein shows that it has autophosphorylation activity. J Bacteriol. 1990 Dec;172(12):6661–6668. doi: 10.1128/jb.172.12.6661-6668.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Parkinson J. S., Kofoid E. C. Communication modules in bacterial signaling proteins. Annu Rev Genet. 1992;26:71–112. doi: 10.1146/annurev.ge.26.120192.000443. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Plamann L., Kuspa A., Kaiser D. Proteins that rescue A-signal-defective mutants of Myxococcus xanthus. J Bacteriol. 1992 May;174(10):3311–3318. doi: 10.1128/jb.174.10.3311-3318.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Plamann L., Li Y., Cantwell B., Mayor J. The Myxococcus xanthus asgA gene encodes a novel signal transduction protein required for multicellular development. J Bacteriol. 1995 Apr;177(8):2014–2020. doi: 10.1128/jb.177.8.2014-2020.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Schröder I., Wolin C. D., Cavicchioli R., Gunsalus R. P. Phosphorylation and dephosphorylation of the NarQ, NarX, and NarL proteins of the nitrate-dependent two-component regulatory system of Escherichia coli. J Bacteriol. 1994 Aug;176(16):4985–4992. doi: 10.1128/jb.176.16.4985-4992.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Stock J. B., Ninfa A. J., Stock A. M. Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev. 1989 Dec;53(4):450–490. doi: 10.1128/mr.53.4.450-490.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Weiss V., Magasanik B. Phosphorylation of nitrogen regulator I (NRI) of Escherichia coli. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8919–8923. doi: 10.1073/pnas.85.23.8919. [DOI] [PMC free article] [PubMed] [Google Scholar]

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