Wednesday, September 3, 2014
Validate protein-protein (and protein - small molecule) interactions in vivo!
So you pulled down your protein complex and high resolution accurate mass spectrometry has identified all of the protein interactors. You follow up with pathway analysis and it all makes sense. All you have to do now is right it up and submit it to Science, right?
Thats probably how it ought to work, right? Unfortunately, not everyone thinks that way, and we are often left "validating" these findings with mixtures of mouse and rabbit blood. Even these old fashioned (outdated...?) techniques have trouble backing up in vivo interactions.
You know what you need? KISS.
No, not these old weirdos...
This KISS: KInase Substrate Sensors, a new technique described in this paper from Brian Chait's lab.
In KISS, multiple antibodies an fluorescent tags can be used to validate that the proteins you said were interacting in vivo really are. Best of all? Metabolite X that you said was involved can be proven to be there as well.
Need to validate something soon? Check this out!