Top-down identification of protein biomarkers in bacteria with unsequenced genomes.
MALDI mass spectrometry-based systems for rapid characterization of microorganisms in biodefense or medical diagnostics usually detect intact proteins in the 5000-20,000 Da range. To evaluate the reliability of species discrimination, and also for forensic applications, it is important that these biomarker proteins be identified. In the present study we apply high resolution tandem mass analysis on an Orbitrap and top-down bioinformatics to identify major biomarker proteins observed in MALDI spectra of intact bacteria for which little genomic or protein sequence information is available. The strategy depends on recognition of proteins with very high homology in related (sequenced) species, making it possible to place unsequenced organisms in their correct phylogenetic context. We show that this rapid proteomics based approach to phylogenetic characterization produces similar results to the traditional techniques, and may even be applied to target organisms of undetermined taxonomy. We further discuss important issues in combining genomics/proteomics databases and MALDI MS for the rapid characterization of microorganisms.
Wynne
Colin
C
Department of Chemistry & Biochemistry, University of Maryland, College Park, Maryland, USA.
Fenselau
Catherine
C
Demirev
Plamen A
PA
Edwards
Nathan
N
D000595:Amino Acid Sequence; D001419:Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry; D015415:Biomarkers / Q000032:analysis; D030562:Databases, Protein; D005240:Feasibility Studies; D016680:Genome, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D040901:Proteomics; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D015007:Yersinia / Q000235:genetics / Q000302:isolation & purification
D001426:Bacterial Proteins; D015415:Biomarkers