Focussed Searches
LUMIER assays allow searches for interaction partners in focussed subsets of the human protome. To this end, collections of expression constructs are generated for selected proteins, e.g. from a specific cellular compartment or pathway. The proteins of the array are expressed with a tag for affinity purification. Such arrayed collections can be tested for interactions with a protein of interest bearing luciferase as a tag.
Compartment-specific subsets
The first collection of clones for array LUMIER we generated is a set of 920 proteins from the nucleus. Test screens have been done with this collection, and have identified novel protein-protein interactions that can be reproduced in classical co-precipitation experiments (immunoprecipitation protocols followed by western blot, unpublished data).
Pathway-specific subsets
We intend to generate small subsets of proteins representing specific signal transduction pathways, such as MAP-kinase signalling, or Wnt-signalling. Interested? Contact us at Frank Schwarz
Click here for a list of currently available expression clones for array LUMIER. These clones are expressed in fusion with the protein A tag for affinity purification. Note that these plates will not sent to customers outside the DKFZ.
Example of an array LUMIER experiment
The picture below shows the result of an array LUMIER experiment. The luciferase-tagged protein in this experiment is CENPT, a component of the centromeric chromatin complex. The protein was tested against a panel of 80 proteins involved in chromatin modulation. Y axis, luciferase activity recovered after purification of the protein A-tagged proteins (arbitrary units). The array contains several fragments of CENPT which gave a strong LUMIER signals, indicating that the protein binds to itsself, be it directly or indirectly.
