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Journal of Bacteriology, July 2003, p. 4268-4275, Vol. 185, No. 14
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.14.4268-4275.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detecting Protein-Protein Interactions in the Intact Cell of Bacillus subtilis (ATCC 6633)

Michael S. Winters{dagger} and R. A. Day*

Department of Chemistry, University of Cincinnati, Cincinnati, Ohio 45221-0172

Received 3 February 2003/ Accepted 28 April 2003

The salt bridge, paired group-specific reagent cyanogen (ethanedinitrile; C2N2) converts naturally occurring pairs of functional groups into covalently linked products. Cyanogen readily permeates cell walls and membranes. When the paired groups are shared between associated proteins, isolation of the covalently linked proteins allows their identity to be assigned. Examination of organisms of known genome sequence permits identification of the linked proteins by mass spectrometric techniques applied to peptides derived from them. The cyanogen-linked proteins were isolated by polyacrylamide gel electrophoresis. Digestion of the isolated proteins with proteases of known specificity afforded sets of peptides that could be analyzed by mass spectrometry. These data were compared with those derived theoretically from the Swiss Protein Database by computer-based comparisons (Protein Prospector; http://prospector.ucsf.edu). Identification of associated proteins in the ribosome of Bacillus subtilis strain ATCC 6633 showed that there is an association homology with the association patterns of the ribosomal proteins of Haloarcula marismortui and Thermus thermophilus. In addition, other proteins involved in protein biosynthesis were shown to be associated with ribosomal proteins.


* Corresponding author. Mailing address: Department of Chemistry, University of Cincinnati, Cincinnati, OH 45221-0172. Phone: (513) 556-9280. Fax: (513) 556-9239. E-mail: Richard.Day{at}Uc.edu.

{dagger} Present address: Protein Chemistry Laboratory, University of Pennsylvania, Philadelphia, PA 19104.


Journal of Bacteriology, July 2003, p. 4268-4275, Vol. 185, No. 14
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.14.4268-4275.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.




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