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Applied and Environmental Microbiology, November 2003, p. 6841-6847, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6841-6847.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Peptide Ligands That Bind Selectively to Spores of Bacillus subtilis and Closely Related Species

Jordan Knurr, Orsolya Benedek,{dagger} Jennifer Heslop, Robert B. Vinson, Jeremy A. Boydston, Joanne McAndrew, John F. Kearney, and Charles L. Turnbough Jr.*

Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294

Received 27 February 2003/ Accepted 16 August 2003

As part of an effort to develop detectors for selected species of bacterial spores, we screened phage display peptide libraries for 7- and 12-mer peptides that bind tightly to spores of Bacillus subtilis. All of the peptides isolated contained the sequence Asn-His-Phe-Leu at the amino terminus and exhibited clear preferences for other amino acids, especially Pro, at positions 5 to 7. We demonstrated that the sequence Asn-His-Phe-Leu-Pro (but not Asn-His-Phe-Leu) was sufficient for tight spore binding. We observed equal 7-mer peptide binding to spores of B. subtilis and its most closely related species, Bacillus amyloliquefaciens, and slightly weaker binding to spores of the closely related species Bacillus globigii. These three species comprise one branch on the Bacillus phylogenetic tree. We did not detect peptide binding to spores of several Bacillus species located on adjacent and nearby branches of the phylogenetic tree nor to vegetative cells of B. subtilis. The sequence Asn-His-Phe-Leu-Pro was used to identify B. subtilis proteins that may employ this peptide for docking to the outer surface of the forespore during spore coat assembly and/or maturation. One such protein, SpsC, appears to be involved in the synthesis of polysaccharide on the spore coat. SpsC contains the Asn-His-Phe-Leu-Pro sequence at positions 6 to 10, and the first five residues of SpsC apparently must be removed to allow spore binding. Finally, we discuss the use of peptide ligands for bacterial detection and the use of short peptide sequences for targeting proteins during spore formation.

* Corresponding author. Mailing address: Department of Microbiology, University of Alabama at Birmingham, BBRB 409, 1530 3rd Ave. South, Birmingham, AL 35294-2170. Phone: (205) 934-6289. Fax: (205) 975-5479. E-mail: ChuckT{at}uab.edu.

{dagger} Present address: Department of Medical Microbiology and Immunology, Faculty of Medicine, University of Pécs, Pécs 7624, Hungary.

Applied and Environmental Microbiology, November 2003, p. 6841-6847, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6841-6847.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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