Working groups

WORKINGGROUP 3

Participants:

Participant	Organization	E-mail	Phone number	Fax
Jiri Stulik, Chair	University of Defence	jstulik@pmfhk.cz	++420495518833	++420495512451
Ludovit Skultety	Institute of Virology	viruludo@savba.sk	+421-2-59302418	+421-2-54774284
Natascha Helena Beyer	Center for Biological Defense	nat@ssi.dk	3268 8403, 3268 8325	3268 3876
Rudolf Toman	Institute of Virology	virutoma@savba.sk	+421-2-59302418	+421-2-54774284
Veljko Veljkovic	Institute of Nuclear Science VINCA	vv@vin.bg.ac.yu	+381 11 2453 686	+38111453686

Description:
WG3 Proteomics and glycomics
- Objective:
  To identify unique proteins as well as glycans from microorganisms as candidate molecules for use in detection/diagnosis, therapy and prophylaxis.
- Description:
  This WG will provide information on unique proteins and glycans that can be used for unambiguous determination of microorganisms and for the identification of targets for diagnosis, antimicrobial therapy and/or vaccine development. Immunogenicity and uniqueness are key factors in determining suitability of candidate molecules towards exclusive recognition of a microbial, viral or fungal species. In addition, this knowledge is indispensable for new leads in antimicrobial therapy and vaccine development. To meet these goals, both proteomic and glycomic approaches will be used and the main focus will be on identification of immunogenic, species and/or strain specific proteins as well as lipopolysaccharides (LPSs).
  Immunogenic proteins derived from the various pathogens will be detected by performing separation of the proteins of a microorganism by means of 2D-PAGE, followed by western blotting with available human or animal sera that contain antibodies that were produced in the course of natural infection. Differential analysis using classical and shotgun proteomics tools followed by image analysis for comparison of protein maps will discover species and/or strain specific proteins. There will be a focus on membrane proteins, cytosolic (sub)fractions and secreted proteins. As glycosylation of bacterial proteins may also play a role in creating uniqueness, attention will also be paid to this post-translational modification.
  To identify proteins of interest that emerge form these investigations, equipment such as MALDI-TOF with PSD/LIFT and LC-ESI-MS/MS, and access to bioinformatics for sequence databases will be necessary. For analysis of functional domains and antigenic determinants the informational spectrum method, a virtual spectroscopy method for structure-function analysis of proteins is available. This information will be used for the design and synthesis of peptides with immunological reactivity.
  As LPSs of pathogenic microorganisms have been considered to be major determinants of virulence expression and infection, and one of the main targets of the antibodies of the host immune system, attention will be paid to structural characterization of LPSs and the subsequent identification, localization, and characterization of their immunoreactive and/or species specific epitopes by combination of chemical and immunological methods. The next step will include characterization and sequencing of the selected, potential diagnostic poly- and oligosaccharides and their modified derivatives, obtained from native LPSs by a number of separation techniques, in GLC-, MALDI-TOF- and EIS-MS.
  The output of these studies should provide complete information on protein topology, amino acid sequence, post-translational modification, presence of functional domains, structural features of LPSs, and composition of immunoreactive epitopes in both proteins and LPSs.

Actions:

Action	Description	download
STSM	The one week stay of two PhD students from Institute of Virology Ing. K. Slaba and Ing. E. Bereghazyov? in proteomic laboratory of Institute of Molecular Pathology, University of Defence .

Publications:

Name	Year	Title	Periodical	Volume	Page	Pubmed
Lenco Juraj	2007	Proteomics analysis of the Francisella tularensis LVS response to iron restriction: Induction of the F. tularensis pathogeinicity island proteins IglABC	FEMS Microbiol Lett	269	11-21
S. Janovska	2007	Identification of immunoreactive antigens in membrane proteins enriched fraction from Francisella tularensis LVS	Immunology Lett.	108	151-159
S. Svraka	2006	Establishment of genotyping scheme of Coxiella burnetii	FEMS Microbiology Letters	254	268-274
I. Pavkova	2006	Comparative proteome analysis of fractions enriched for membrane-associated proteins from Francisella tularensis subsp. tularensis and F. tularensis subsp. holarctica strains.	Journal of Proteome Research	11	3125-34
Jana Havlasov	2005	Proteomic analysis of anti-Francisella tularensis LVS antibody response in murine model of tularemia	Proteomics	5	2090-103
Roman Hrstka	2005	The role of MAPK signal pathways during Francisella tularensis LVS infection-induced apoptosis in murine macrophages	Microbes and Infection	7	619-625
Juraj Lenco	2005	. Insights into the oxidative stress response in Francisella tularensis LVS and its mutant DeltaiglC1+2 by proteomics analysis	FEMS Microbiology Letters	246	47-54
Ivona P?vkov	2005	Francisella tularensis live vaccine strain: proteomic analysis of membrane proteins enriched fraction.	Proteomics	5	2460-2467
P. Vadovic	2005	Structural and functional characterization of the glycan antigens involved in immunobiology of Q fever	ANNALS OF THE NEW YORK ACADEMY OF SCIENCES	1063	149-153
M. Fodorova	2005	Structural features of lipopolysaccharide from Rickettsia typhi, the causative agent of endemic typhus	ANNALS OF THE NEW YORK ACADEMY OF SCIENCES	1063	259-260
L. Skultety	2005	Coxiella burnetii whole cell lysate protein identification by mass spectrometry and	ANNALS OF THE NEW YORK ACADEMY OF SCIENCES	1063	115-122
E. Kov?čov	2005	Analysis of specificity of interaction between synthetic key saccharide components of lipopolysaccharides and monoclonal antibodies to Coxiella burnetii	ACTA VIROLOGICA	49	261-270