FULL CHARACTERIZATION OF NATURAL KILLER CELL MEMBRANE
MICRODOMAINS
P. Man1,2, P. Novák1,2, P. Pompach1,2,
D. Ulbrichová1,2, V. Havlíček1, K. Bezouška1,2
1Institute of Microbiology, Academy of Sciences
of the Czech Republic, Prague, CZ
2Department of Biochemistry, Faculty of Science,
Charles University, Prague, CZ
Natural
killer (NK) cells are cytotoxic effector lymphocytes, which do not express
antigen-specific cell surface receptors. NK receptors that mediate signals
leading to the initiation or supression of natural cytotoxicity processes are
poorly characterized. Moreover, these receptors may associate with other
co-stimulating molecules and adaptor proteins, which transduce the signal from
the receptor to the cell. Such complexes may be observed as biochemically
distinct parts of plasmatic membranes and are commonly referred to micro domains
or glycosphingolipid enriched microdomains (GEMs). They are enriched in
GPI-anchored and acetylated proteins and in cholesterol and glycosphingolipids.
It is believed that they may aid in signal transduction as well as in
trafficking through the secretory and endocytic pathways and in cell to cell interaction.
In
this study we focused on membrane microdomains from rat NK leukaemia cell line
(RNK-16). For detailed characterization we used a shotgun strategy based on
microcapillary HPLC – tandem mass spectrometry. Aditionally, we applied
techniques of native electrophoresis for detailed mapping of protein complexes
present in the GEMs. We have identified a large number of proteins (e.g. gp-42,
CD2, LAT, CD161, CD44 or g-proteins in GEM and tubuline in non-GEM fractions).
Financial
support: MSM 113100001, AV0Z5020903.