An enzyme squirted out of deep sea shrimp causing a bright blue burst to scare away predators is the key to innovative scientific work to develop new therapies for serious human diseases.
Associate Professor Kevin Pfleger and colleagues, from the Harry Perkins Institute of Medical Research, The University of Nottingham and Promega Corporation, tested a derivative of the bioluminescent protein and found it was a “game-changer” for monitoring the way hormones and pharmaceuticals bind to receptors on the surface of living cells.
Associate Professor Pfleger, who is Head of Molecular Endocrinology and Pharmacology at the Perkins, says they used an exciting new technology known as NanoBRET developed by Promega, with the latest instrumentation from BMG Labtech Australia.
Associate Professor Pfleger says that understanding how medicines bind to receptors on the cell is incredibly important for discovering and developing new therapies to help patients.
“Our world-first approach enables this binding to be observed in live cells in real-time under physiological conditions in a way that has not been possible before”, he says.
The key to future breakthroughs is through understanding the nuances of how medicines need to work.
“If cell function is like playing the piano, most medicines slam the keys or close the lid. The idea is to get back to the tune the body is trying to play,” he says.
The types of receptors Associate Professor Pfleger studies are the targets for about a third of the current pharmaceuticals on the Australian market. These include medication for high blood pressure, neurological disorders such as Parkinson’s disease, asthma, pain and inflammation, reproductive disorders, kidney disease and cancer.
This work was funded by an Australian Research Council Linkage Grant and the results have been published in the July edition of the very high impact scientific journal, Nature Methods.
Image courtesy of Promega
A receptor (shown in purple) in the cell membrane is fused to the luminescent NanoLuc enzyme derived from that found in a deep sea shrimp. When a small molecule pharmaceutical attached to a fluorescent molecule binds to the receptor, energy is transferred from the NanoLuc that excites the fluorescent molecule and makes it glow red, like turning on a tiny red fluorescent light bulb.
This red light can be measured in live cells in real time, and is only seen when the fluorescently-labelled pharmaceutical is bound to the receptor, and therefore close enough to the NanoLuc for energy transfer. Unlabelled pharmaceuticals that compete for binding to the receptor can then be tested, their binding inferred by a decrease in red light as the fluorescently-labelled compound is no longer able to bind to the receptor.