Proteins in touch with lipids - DOC by hcj


									Proteins in touch with lipids

Contact person:        Dr. Per Haberkant, Membrane Enzymology
                       phone: +31 30 253 24 58

Lipids are one of the four major molecular components of biological organisms, along
with nucleic acids, proteins and sugars. Lipids are constituents of cellular membranes and
are embedded in numerous signaling events. Derailed lipid homeostasis contributes to the
pathogenesis of human diseases as for example cancer, diabetes and Alzheimer’s disease.
In order to understand how lipids function within a cell one has to identify the keyplayers
in the life of individual lipids.
We recently developed a new technique that enables to study protein-lipid interactions in
vivo. The method is based on bifunctional lipids that contain (i) a photoactivatable group
and (ii) a “clickable” moiety. In order to identify proteins interacting with lipids we use a
“two-click strategy”: First, proteins interacting with bifunctional lipids can be tagged by
the lipid itself, applying photocrosslinking. Second, subsequent tagging of the
photoaffinity labeled protein by means of click chemistry will allow
purification/visualization of crosslinked products.
The technique is currently used to identify proteins with a high affinity for individual
lipids. This will give new insights in how lipids are embedded in cellular functions and
will open new perspectives for therapeutic intervention of lipid-regulated processes in a
number of pathological situations.


Synthesis of bifunctional lipids, cell culture, photoaffinity labeling, purification of
crosslinked products, analysis of crosslinked products by SDS-PAGE, Western blotting,
in-gel fluorescence, mass spectrometry

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