Journal article
Biomolecular interaction analysis in functional proteomics



Publication Details
Authors:
Bertinetti, D.; Prinz, A.; Gesellchen, F.; Herberg, F.; Zimmermann, B.; Drewianka, S.
Publication year:
2006
Journal:
Journal of Neural Transmission
Pages range:
1015-1032
Volume number:
113
Start page:
1015
End page:
1032
ISSN:
0300-9564

Abstract
To understand the function of highly complex eukaryotic tissues like the human brain, in depth knowledge about cellular protein networks is required. Biomolecular interaction analysis (BIA), as a part of functional proteomics, aims to quantify interaction patterns within a protein network in detail. We used the cAMP dependent protein kinase (PKA) as a model system for the binding analysis between small natural ligands, cAMP and cAMP analogues, with their physiological interaction partner, the regulatory subunit of PKA. BIA comprises a variety of methods based on physics, biochemistry and molecular biology. Here we compared side by side real time SPR (surface plasmon resonance, Biacore), a bead based assay (AlphaScreen), a fluorescence based method (Fluorescence polarisation) and ITC (isothermal titration calorimetry). These in vitro methods were complemented by an in cell reporter assay, BRET2 (bioluminescence resonance energy transfer), allowing to test the effects of cAMP analogues in living cells.

Last updated on 2019-25-07 at 10:42