Quantotypic Properties of QconCAT Peptides Targeting Bovine Host Response to Streptococcus uberis

• Published in: Journal of proteome research 2012-03, Vol.11 (3), p.1832-1843
• Main Authors: Bislev, Stine L, Kusebauch, Ulrike, Codrea, Marius C, Beynon, Robert J, Harman, Victoria M, Røntved, Christine M, Aebersold, Ruedi, Moritz, Robert L, Bendixen, Emøke
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Cattle; Chromatography, Ion Exchange - standards; Female; Host-Pathogen Interactions; Immunologic Factors; Inflammation Mediators - chemistry; Inflammation Mediators - isolation & purification; Inflammation Mediators - metabolism; Mammary Glands, Animal - metabolism; Mammary Glands, Animal - microbiology; Mass Spectrometry - standards; Mastitis, Bovine - metabolism; Mastitis, Bovine - microbiology; Molecular Sequence Data; Peptide Fragments - chemistry; Peptide Mapping - methods; Peptide Mapping - standards; Protein Stability; Proteolysis; Proteome - chemistry; Proteome - isolation & purification; Proteome - metabolism; Reference Standards; Reproducibility of Results; Streptococcal Infections - metabolism; Streptococcal Infections - microbiology; Streptococcal Infections - veterinary; Streptococcus - physiology
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/pr201064g

Mammalian host response to pathogens is associated with fluctuations in high abundant proteins in body fluids as well as in regulation of proteins expressed in relatively low copy numbers like cytokines secreted from immune cells and endothelium. Hence, efficient monitoring of proteins associated with host response to pathogens remains a challenging task. In this paper, we present a targeted proteome analysis of a panel of 20 proteins that are widely believed to be key players and indicators of bovine host response to mastitis pathogens. Stable isotope-labeled variants of two concordant proteotypic peptides from each of these 20 proteins were obtained through the QconCAT method. We present the quantotypic properties of these 40 proteotypic peptides and discuss their application to research in host–pathogen interactions. Our results clearly demonstrate a robust monitoring of 17 targeted host-response proteins. Twelve of these were readily quantified in a simple extraction of mammary gland tissues, while the expression levels of the remaining proteins were too low for direct and stable quantification; hence, their accurate quantification requires further fractionation of mammary gland tissues.