Quantotypic properties of QconCAT peptides targeting bovine host response to Streptococcus uberis.

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.

[1]  R. Zadoks,et al.  Host-response patterns of intramammary infections in dairy cows. , 2011, Veterinary immunology and immunopathology.

[2]  Carolyn L. Geczy,et al.  Inflammation-associated S100 proteins: new mechanisms that regulate function , 2011, Amino Acids.

[3]  F. Lottspeich,et al.  Evaluation of protein loading techniques and improved separation in OFFGEL isoelectric focusing , 2011, Electrophoresis.

[4]  Jeffrey DeGrasse,et al.  The proteomic advantage: label-free quantification of proteins expressed in bovine milk during experimentally induced coliform mastitis. , 2010, Veterinary immunology and immunopathology.

[5]  Maren S Fragala,et al.  Conceptual and methodological issues relevant to cytokine and inflammatory marker measurements in clinical research , 2010, Current opinion in clinical nutrition and metabolic care.

[6]  E. Bendixen,et al.  Quantitative milk proteomics – Host responses to lipopolysaccharide‐mediated inflammation of bovine mammary gland , 2010, Proteomics.

[7]  N. Trivedi,et al.  Mast cell peptidases: chameleons of innate immunity and host defense. , 2010, American journal of respiratory cell and molecular biology.

[8]  M. McFarland,et al.  Proteomic analysis of the temporal expression of bovine milk proteins during coliform mastitis and label-free relative quantification. , 2010, Journal of dairy science.

[9]  R. Aebersold,et al.  Perspectives of targeted mass spectrometry for protein biomarker verification. , 2009, Current opinion in chemical biology.

[10]  R. Aebersold,et al.  Applying mass spectrometry-based proteomics to genetics, genomics and network biology , 2009, Nature Reviews Genetics.

[11]  Lukas N. Mueller,et al.  Full Dynamic Range Proteome Analysis of S. cerevisiae by Targeted Proteomics , 2009, Cell.

[12]  Richard O'Kennedy,et al.  Mastitis detection: current trends and future perspectives. , 2009, Trends in biotechnology.

[13]  D. Bannerman.  Pathogen-dependent induction of cytokines and other soluble inflammatory mediators during intramammary infection of dairy cows. , 2009, Journal of animal science.

[14]  J. Lunney,et al.  A current perspective on availability of tools, resources and networks for veterinary immunology. , 2009, Veterinary immunology and immunopathology.

[15]  Ruedi Aebersold,et al.  Targeted proteomic strategy for clinical biomarker discovery , 2009, Molecular oncology.

[16]  D. Bannerman,et al.  Proteomic analysis of differentially expressed proteins in bovine milk during experimentally induced Escherichia coli mastitis. , 2008, Journal of dairy science.

[17]  R. Aebersold,et al.  Selected reaction monitoring for quantitative proteomics: a tutorial , 2008, Molecular systems biology.

[18]  B. Guldbrandtsen,et al.  Detection of quantitative trait loci in Danish Holstein cattle affecting clinical mastitis, somatic cell score, udder conformation traits, and assessment of associated effects on milk yield. , 2008, Journal of dairy science.

[19]  A. Hokama,et al.  Roles of galectins in inflammatory bowel disease. , 2008, World journal of gastroenterology.

[20]  Nichole L. King,et al.  Targeted Quantitative Analysis of Streptococcus pyogenes Virulence Factors by Multiple Reaction Monitoring*S , 2008, Molecular & Cellular Proteomics.

[21]  B. Guldbrandtsen,et al.  Pathogen-specific effects of quantitative trait loci affecting clinical mastitis and somatic cell count in Danish Holstein cattle. , 2008, Journal of dairy science.

[22]  Christine A. Miller,et al.  Efficient Fractionation and Improved Protein Identification by Peptide OFFGEL Electrophoresis*S , 2006, Molecular & Cellular Proteomics.

[23]  R. Beynon,et al.  Multiplexed absolute quantification for proteomics using concatenated signature peptides encoded by QconCAT genes , 2006, Nature Protocols.

[24]  P. Rainard,et al.  Innate immunity of the bovine mammary gland. , 2006, Veterinary research.

[25]  R. Beynon,et al.  Multiplexed absolute quantification in proteomics using artificial QCAT proteins of concatenated signature peptides , 2005, Nature Methods.

[26]  C. Røntved,et al.  Effects of diet energy density and milking frequency in early lactation on tumor necrosis factor-alpha responsiveness in dairy cows. , 2005, Veterinary immunology and immunopathology.

[27]  Richard A. Young,et al.  Insights into host responses against pathogens from transcriptional profiling , 2005, Nature Reviews Microbiology.

[28]  M. Paape,et al.  Innate immune response to intramammary infection with Serratia marcescens and Streptococcus uberis. , 2004, Veterinary research.

[29]  M. Paape,et al.  Escherichia coli and Staphylococcus aureus Elicit Differential Innate Immune Responses following Intramammary Infection , 2004, Clinical Diagnostic Laboratory Immunology.

[30]  M. Paape,et al.  Increased levels of LPS-binding protein in bovine blood and milk following bacterial lipopolysaccharide challenge. , 2003, Journal of dairy science.

[31]  Christine Fourichon,et al.  Production effects related to mastitis and mastitis economics in dairy cattle herds. , 2003, Veterinary research.

[32]  Satu Pyörälä,et al.  Indicators of inflammation in the diagnosis of mastitis. , 2003, Veterinary research.

[33]  P. Heegaard,et al.  Early pathogenesis and inflammatory response in experimental bovine mastitis due to Streptococcus uberis. , 2003, Journal of comparative pathology.

[34]  J. Bienvenu,et al.  Cytokine assays in human sera and tissues. , 1998, Toxicology.

[35]  R. Aebersold,et al.  High-throughput generation of selected reaction-monitoring assays for proteins and proteomes , 2010, Nature Methods.

[36]  S. Davis,et al.  Transcriptome profiling of Streptococcus uberis-induced mastitis reveals fundamental differences between immune gene expression in the mammary gland and in a primary cell culture model. , 2009, Journal of dairy science.

[37]  Daniel B. Martin,et al.  Computational prediction of proteotypic peptides for quantitative proteomics , 2007, Nature Biotechnology.