title: Protein markers for identification of Yersinia pestis and their variation related to culture.
abstract: The detection of high consequence pathogens, such as Yersinia pestis, is well established in biodefense laboratories for bioterror situations. Laboratory protocols are well established using specified culture media and a growth temperature of 37 °C for expression of specific antigens. Direct detection of Y. pestis protein markers, without prior culture, depends on their expression. Unfortunately protein expression can be impacted by the culture medium which cannot be predicted ahead of time. Furthermore, higher biomass yields are obtained at the optimal growth temperature (i.e. 28 °C-30 °C) and therefore are more likely to be used for bulk production. Analysis of Y. pestis grown on several types of media at 30 °C showed that several protein markers were found to be differentially detected in different media. Analysis of the identified proteins against a comprehensive database provided an additional level of organism identification. Peptides corresponding to variable regions of some proteins could separate large groups of strains and aid in organism identification. This work illustrates the need to understand variability of protein expression for detection targets. The potential for relating expression changes of known proteins to specific media factors, even in nutrient rich and chemically complex culture medium, may provide the opportunity to draw forensic information from protein profiles.
authors: Pacific Northwest National Laboratory, Richland, WA 99352, USA. Electronic address: David.Wunschel@pnnl.gov.
authors: David
authors: D
authors: Wunschel
authors: Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors: Heather
authors: H
authors: Engelmann
authors: Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors: Kristin
authors: K
authors: Victry
authors: Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors: Brian
authors: B
authors: Clowers
authors: Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors: Christina
authors: C
authors: Sorensen
authors: Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors: Nancy
authors: N
authors: Valentine
authors: Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors: Christine
authors: C
authors: Mahoney
authors: Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors: Thomas
authors: T
authors: Wietsma
authors: Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors: Karen
authors: K
authors: Wahl
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media / Q000737:chemistry*; D030562:Databases, Protein; D015964:Gene Expression Regulation, Bacterial; D014644:Genetic Variation; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D053719:Tandem Mass Spectrometry; D015010:Yersinia pestis / Q000145:classification / Q000254:growth & development*
chemical_list: D001426:Bacterial Proteins; D015415:Biomarkers; D003470:Culture Media; D010455:Peptides
keywords: Culture environment; HPLC; Protein expression; Tandem mass spectrometry; Yersinia pestis