Discovery and Targeted Proteomics on Cutaneous Biopsies Infected by Borrelia to Investigate Lyme Disease*
Electrophoresis
Proteomics
0301 basic medicine
Antigens, Bacterial
Lyme Disease
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
DOI:
10.1074/mcp.m114.046540
Publication Date:
2015-02-26T13:33:23Z
AUTHORS (8)
ABSTRACT
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).
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