MICROPLATE READERS
Protein-protein
INTERACTIONS P
Dr Barry Whyte discusses how microplate readers could help accelerate drug discovery and development
rotein-protein interactions, which underpin many crucial molecular events taking place in a cell, are
an important area of research and discovery. A typical cell contains thousands of proteins and few of them function alone. It is therefore vital to study how proteins interact with one another and the tools to investigate these interactions have continued to advance over time. Applications amenable to scale can signifi cantly accelerate research and discovery of inhibitors and drugs, for instance. Here, we look at a few examples of how detection technologies in microplate readers provide options for the study of protein-protein interactions in the rapidly emerging area of targeted protein degradation.
DIFFERENT OPTIONS TO STUDY PROTEIN- PROTEIN INTERACTIONS Microplate readers support many
24
www.scientistive.com
useful applications in the life sciences due to their ability to provide ready access to a range of detection technologies. Protein-protein interactions require eff icient, highly- sensitive assays often with many measurements in a short period of time. For the screening of molecules, it is crucial to have detection technologies that are compatible with automation, and which deliver the required sensitivity for miniaturised assays. Fluorescence- and luminescence-based measurements are often used for this purpose. In addition to detection methods based on Förster’s Resonance Energy Transfer (FRET), fl uorescence polarisation is an advanced detection mode that off ers exciting opportunities for the
study of protein-protein interactions. ATTECs (Autophagy-Tethering
Compounds) are a novel class of bifunctional molecules proposed to hijack the autophagosomal pathway for the degradation of cellular components including proteins. The induction of Atg8 family protein interactions with target proteins of interest off ers the possibility for novel targeted protein degradation approaches. LC3A is a member of the human Atg8 protein family and is only found in the autophagosome. The discovery of potent LC3A inhibitors would therefore serve as a handle for the development of ATTECs.1 In the following example, a
fl uorescence polarisation assay was developed based on the use of a
Figure 1: Adding competing ligands of LC3A to the LC3A-tracer complex results in a displacement of tracer
Page 1 |
Page 2 |
Page 3 |
Page 4 |
Page 5 |
Page 6 |
Page 7 |
Page 8 |
Page 9 |
Page 10 |
Page 11 |
Page 12 |
Page 13 |
Page 14 |
Page 15 |
Page 16 |
Page 17 |
Page 18 |
Page 19 |
Page 20 |
Page 21 |
Page 22 |
Page 23 |
Page 24 |
Page 25 |
Page 26 |
Page 27 |
Page 28 |
Page 29 |
Page 30 |
Page 31 |
Page 32 |
Page 33 |
Page 34 |
Page 35 |
Page 36 |
Page 37 |
Page 38 |
Page 39 |
Page 40 |
Page 41 |
Page 42 |
Page 43 |
Page 44 |
Page 45 |
Page 46 |
Page 47 |
Page 48 |
Page 49 |
Page 50 |
Page 51 |
Page 52 |
Page 53 |
Page 54 |
Page 55 |
Page 56 |
Page 57 |
Page 58 |
Page 59 |
Page 60
