This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Herrmann, S.
Right arrow Articles by Taylor, B. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Herrmann, S.
Right arrow Articles by Taylor, B. L.

 Previous Article  |  Next Article 

Journal of Bacteriology, October 2004, p. 6782-6791, Vol. 186, No. 20
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.20.6782-6791.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

PAS Domain of the Aer Redox Sensor Requires C-Terminal Residues for Native-Fold Formation and Flavin Adenine Dinucleotide Binding

Sarah Herrmann, Qinhong Ma,{dagger} Mark S. Johnson, Alexandre V. Repik,{ddagger} and Barry L. Taylor*

Division of Microbiology and Molecular Genetics, Loma Linda University, Loma Linda, California

Received 17 May 2004/ Accepted 19 July 2004

The Aer protein in Escherichia coli is a membrane-bound, FAD-containing aerotaxis and energy sensor that putatively monitors the redox state of the electron transport system. Binding of FAD to Aer requires the N-terminal PAS domain and residues in the F1 region and C-terminal HAMP domain. The PAS domains of other PAS proteins are soluble in water. To investigate properties of the PAS domain, we subcloned segments of the aer gene from E. coli that encode the PAS domain with and without His6 tags and expressed the PAS peptides in E. coli. The 20-kDa His6-Aer2-166 PAS-F1 fragment was purified as an 800-kDa complex by gel filtration chromatography, and the associating protein was identified by N-terminal sequencing as the chaperone protein GroEL. None of the N-terminal fragments of Aer found in the soluble fraction was released from GroEL, suggesting that these peptides do not fold correctly in an aqueous environment and require a motif external to the PAS domain for proper folding. Consistent with this model, peptide fragments that included the membrane binding region and part (Aer2-231) or all (Aer2-285) of the HAMP domain inserted into the membrane, indicating that they were released by GroEL. Aer2-285, but not Aer2-231, bound FAD, confirming the requirement for the HAMP domain in stabilizing FAD binding. The results raise an interesting possibility that residues outside the PAS domain that are required for FAD binding are essential for formation of the PAS native fold.

* Corresponding author. Mailing address: Division of Microbiology and Molecular Genetics, Loma Linda University, Loma Linda, CA 92350. Phone: (909) 558-8544. Fax: (909) 558-0244. E-mail: bltaylor{at}univ.llu.edu.

{dagger} Present address: Dow Chemical Co., San Diego, CA 92122.

{ddagger} Present address: University of Massachusetts Medical School, Worcester, MA 01605-2324.

Journal of Bacteriology, October 2004, p. 6782-6791, Vol. 186, No. 20
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.20.6782-6791.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Watts, K. J., Johnson, M. S., Taylor, B. L. (2008). Structure-Function Relationships in the HAMP and Proximal Signaling Domains of the Aerotaxis Receptor Aer. J. Bacteriol. 190: 2118-2127 [Abstract] [Full Text]  
  • Amin, D. N., Taylor, B. L., Johnson, M. S. (2007). Organization of the Aerotaxis Receptor Aer in the Membrane of Escherichia coli. J. Bacteriol. 189: 7206-7212 [Abstract] [Full Text]  
  • del Carmen Buron-Barral, M., Gosink, K. K., Parkinson, J. S. (2006). Loss- and Gain-of-Function Mutations in the F1-HAMP Region of the Escherichia coli Aerotaxis Transducer Aer.. J. Bacteriol. 188: 3477-3486 [Abstract] [Full Text]  
  • Watts, K. J., Sommer, K., Fry, S. L., Johnson, M. S., Taylor, B. L. (2006). Function of the N-Terminal Cap of the PAS Domain in Signaling by the Aerotaxis Receptor Aer. J. Bacteriol. 188: 2154-2162 [Abstract] [Full Text]  
  • Amin, D. N., Taylor, B. L., Johnson, M. S. (2006). Topology and Boundaries of the Aerotaxis Receptor Aer in the Membrane of Escherichia coli. J. Bacteriol. 188: 894-901 [Abstract] [Full Text]  
  • Aranda-Olmedo, I., Ramos, J. L., Marques, S. (2005). Integration of Signals through Crc and PtsN in Catabolite Repression of Pseudomonas putida TOL Plasmid pWW0. Appl. Environ. Microbiol. 71: 4191-4198 [Abstract] [Full Text]  
  • Hong, C. S., Kuroda, A., Takiguchi, N., Ohtake, H., Kato, J. (2005). Expression of Pseudomonas aeruginosa aer-2, One of Two Aerotaxis Transducer Genes, Is Controlled by RpoS. J. Bacteriol. 187: 1533-1535 [Abstract] [Full Text]  
  • Ma, Q., Johnson, M. S., Taylor, B. L. (2005). Genetic Analysis of the HAMP Domain of the Aer Aerotaxis Sensor Localizes Flavin Adenine Dinucleotide-Binding Determinants to the AS-2 Helix. J. Bacteriol. 187: 193-201 [Abstract] [Full Text]  
  • Ma, Q., Roy, F., Herrmann, S., Taylor, B. L., Johnson, M. S. (2004). The Aer Protein of Escherichia coli Forms a Homodimer Independent of the Signaling Domain and Flavin Adenine Dinucleotide Binding. J. Bacteriol. 186: 7456-7459 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»