Discovery and Targeted Proteomics on Cutaneous Biopsies Infected by Borrelia to Investigate Lyme Disease*

• DOI: 10.1074/mcp.m114.046540
• Publication Date: 2015-02-26T13:33:23Z
• Authors: Gilles Schnell; Amandine Boeuf; Benoît Westermann; Benoît Jaulhac; Dan Lipsker; Christine Carapito; Nathalie Boulanger; Laurence Ehret-Sa...
• Keywords: Electrophoresis; Proteomics; 0301 basic medicine; Antigens, Bacterial; Lyme Disease; 0303 health sciences; Biopsy; 3. Good health; Mice; 03 medical and health sciences; Borrelia burgdorferi; Isotope Labeling; Animals; Humans; Peptides; Gels; Bacterial Outer Membrane Proteins; Chromatography, Liquid; Flagellin; Skin

Lyme disease is the most important vector-borne in Northern hemisphere and represents a major public health challenge with insufficient means of reliable diagnosis. Skin rarely investigated proteomics but constitutes case key interface where pathogens can enter, persist, multiply. Therefore, we on skin samples to detect Borrelia proteins directly cutaneous biopsies robust specific way. We first set up discovery gel prefractionation-LC-MS/MS approach murine model infected by burgdorferi sensu stricto that allowed identification 25 among more than 1300 mouse proteins. Then developed targeted prefractionation-LC-selected reaction monitoring (SRM) assay 9/33 proteins/peptides tissue using heavy labeled synthetic peptides. successfully transferred this from human (naturally Borrelia), were able two proteins: OspC flagellin. Considering extreme variability OspC, an extended SRM target large variants. This afforded detection nine peptides belonging either or flagellin biopsies. further shortened sample preparation showed detectable liquid digestion followed LC-SRM analysis without any prefractionation. study thus shows promising tool for early direct diagnosis high sensitivity (<10 fmol OspC/mg biopsy).