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Applied and Environmental Microbiology, February 2003, p. 1100-1107, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.1100-1107.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Small, Acid-Soluble Proteins as Biomarkers in Mass Spectrometry Analysis of Bacillus Spores

Yetrib Hathout,1* Barbara Setlow,2 Rosa-Martha Cabrera-Martinez,2,{dagger} Catherine Fenselau,1 and Peter Setlow2

Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20704,1 Department of Biochemistry, University of Connecticut Health Center, Farmington, Connecticut 060322

Received 5 August 2002/ Accepted 28 October 2002

The use of 1 N HCl for extraction of small, acid-soluble proteins (SASP) from different Bacillus spore species was examined. The extracts were analyzed by high-performance liquid chromatography and matrix-assisted laser desorption mass spectrometry and were found to be both qualitatively and quantitatively superior to extraction by acetonitrile-5% trifluoroacetic acid (70:30, vol/vol). Both major and minor {alpha}/ß- and {gamma}-type SASP were characterized by their molecular masses or tryptic peptide maps and by searches of both protein and unannotated genome databases. For all but 1 pair (B. cereus T and B. thuringiensis subsp. Kurstaki) among the 11 variants studied the suites of SASP masses are distinctive, consistent with the use of these proteins as potential biomarkers for spore identification by mass spectrometry.

* Corresponding author. Mailing address: Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742. Phone: (301) 405-8619. Fax: (301) 405-8615. E-mail: hathout{at}wam.umd.edu.

{dagger} Present address: Departmento de Bioquimica, Escuela Nacional de Ciencas Biologicas, Instituto Politecnico Nacional, Mexico City, Mexico 11340.

Applied and Environmental Microbiology, February 2003, p. 1100-1107, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.1100-1107.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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