StarPATT

We have recently introduced a method that is capable of detecting and quantifying protein-protein-interactions (PPIs) in a live cell context (Schwarzenbacher, Kaltenbrunner et al. 2008; Weghuber, Brameshuber et al. 2010). This approach, termed µ-patterning technique, is based on the combination of fluorescent microscopy and µ-patterned surfaces, and turned out to be a valuable tool to address different biological questions. Most importantly, it offers the possibility to quantify PPIs with high-throughput. In order to exploit the high-throughput capability, further development steps have to be tackled, which we want to do within this project. The addressed questions include:
Ø  Further improvements in experiment design and materials: We have recognized that different biological questions require different pattern sizes. For example smaller cells (e.g. suspension cells) demand for small structures and may improve the adhesion process of the cell. Furthermore, variations in the µ-structures (e.g. patterns of different size as well as lines) enable the enrichment of a ligand at certain positions. Finally, such µ-structure variations enable the performance of functional studies (e.g. analysis of PPIs within focal adhesions of migrating cells or signaling clusters). We will use novel state-of-the-art methodologies to evaluate the capabilities of different stamp materials varying in size and/or nature.

Ø  Analysis of medically relevant PPIs: For the characterization of the stamp materials we will also focus on biological systems with great medical relevance. Such candidate PPIs (e.g. G-protein coupled receptors) would be used for drug screening approaches, thus the evaluation will be aimed at these interaction partners.

Development of a multi-well imaging plate with an optimized µ-structured surface: To tap the full potential of the µ-patterning approach we will develop a prototype imaging plate with a functionalized and µ-structured surface. It will offer users working in different fields of life-sciences the possibility to analyze many PPIs within limited time. Furthermore, the testing of great numbers of active ingredients (no matter if natural of synthetic) on certain PPIs will become possible.