Project information

Integrated genome and proteome analysis of therapeutically important bacteriophages by combination of electrophoresis and mass spectrometry

Investor logo
Project Identification
GA203/03/0515
Project Period
1/2003 - 1/2005
Investor / Pogramme / Project type
Czech Science Foundation
MU Faculty or unit
Faculty of Science
Keywords
mass spectrometry, capillary electrophoresis, proteome, bacteriophage

It is expected that selected bacteriophages (bacterial viruses), or even
their specific proteins and/or peptides can be used as new effective means
to treat bacterial infections. For a rational development of phage therapy,
without the adverse effects of current antibiotics, the relationship between
desired biological properties of suitable phages and their genome/proteome
structure must be found. To achieve this goal it is necessary to find new
technologies for rapid analysis of the phage molecular structure. In this
project, we will develop a three-dimensional separation system for analysis
and typing of the phage proteome. First, micropreparative capillary
electrophoresis with laser-induced fluorescence detection (LIF) and
noncovalent labeling will be used for the separation of the phage proteins.
This fingerprint will be used to trace differences in protein content of
phages and to control the automated fraction collection. The resulting
fractions will be digested by trypsin immobilized directly on the walls of
the collection micro wells. In the final step, the tryptic peptides will be
analyzed by capillary electrophoresis coupled on-line to mass spectrometry.
To provide the peptide sequence and/or information on post-translational
modifications both the electrospray with ion trap (ESI/IT) and matrix
assisted laser desorption/ionization with time-of-flight (MALDI/TOF) mass
spectrometers will be used as complementary tools. Phage 812 and its
host-range mutants, which exhibit strong lytic activity and different
host-specificity, have been chosen for this study.

Publications

Total number of publications: 32


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