XXII. BIOCHEMICKÝ ZJAZD - Jesseniova lekárska fakulta

Lectures
IDENTIfICaTION of SurfaCE PrOTEINS of THE OBLIGaTE
INTraCELLULar baCTErIUM CoXIeLLa burneTII
Gabriela Flores Ramírez 1 , Zuzana Bílková 2 , Pavol Vadovič 1 and Ľudovít Škultéty 1,3
1
Department of Rickettsiology. Institute of Virology, SAS, 845 05 Bratislava, Slovakia.
2
Department of Biological and Biochemical Sciences, Faculty of Chemical Technology,
University of Pardubice, Pardubice, Czech Republic, 3 Centre of Molecular Medicine,
Slovak Academy of Sciences, Bratislava, Slovakia.
Coxiella burnetti is an obligatory, intracellular bacterium that causes Q fever in humans.
Due to the important role of surface proteins in adhesion, invasion, and intracellular
survival of pathogens in the host, these proteins represent the crucial bacterial antigens,
drug targets or biomarkers for diagnostic purposes. The aim of our investigation was to
extract and identify the surface proteins of C. burnetii in virulent phase I.
Two different strategies were applied. The first is based on protein extraction by detergent
Triton X-114 followed by separation of the extracted proteins by SDS-PAGE and tryptic
digestion. The second one is the surface shaving procedure employing magnetic beads
with immobilized trypsin. The tryptic peptides were then separated by nanocapillary liquid
chromatography and the surface proteins were identified by tandem mass spectrometry.
Each of the presented methods has some advantages over those conventional ones and
they allowed us to identify high numbers of proteins predicted to be surface localized. In
addition to those lipoproteins, ribosomal proteins and metabolic enzymes were identified.
Acknowledgements: This contribution/publication is the result of the project implementation:
„TRANSMED“ supported by the Research & Development Operational Programme
funded by the ERDF.
XXII. Biochemistry Congress, Martin
93