Wednesday, October 9, 2019

A proximity biotinylation map of human cells!

This image of how BioID works was brazenly stolen from Creative Biolabs -- sorry, please let me know if you want me to take it down -- because they already offer this BioID service!

I'm using this because the preprint has some strict copyright statements on their pictures, however -- this is worth checking out.

What is it?

Oh -- just a measurement of how 4,000 + proteins hang out with each other. This is a hot new technology (at least, I never heard of it until recently -- okay -- I'm just dumb -- here is a 2013 paper  -- wait -- i think it has become a little more refined over time...)

How's it do it?

You use a "promiscuous" protein ligase that you FUSE to your protein of interest. Then you dump in a bunch of biotin. The end of your protein with the fusion can only cause a biotin connection with things that are in close proximity to it. Apparently people have pulled this off in vivo?

I guess there are some assumptions -- like your fusion doesn't completely negate your protein - protein interactions? Anyway, if it is close then -- BOOM! the biotin gets all ligated and then you enrich your proximity proteins with normal biotin enriching thingies.

Does this mean that this group fused a few thousand proteins? Yeah -- I think it does....?

Does that sound crazy? totally does.

Work was done on an Orbitrap Elite using a high/low (Orbitrap CID-Iontrap method). My assumption is that they bought it right when it came out for this project (ASMS 2012?) and this paper is the summary of what that Orbitrap was doing the last 7 years or so....

All the data is deposited at PRIDE, but I can't find it on this tablet since there are 20 papers in 2019 on this BioID stuff that have deposited data and I'm sleepy. (A lot of times the PRIDE doesn't go live until the paper is accepted anyway)

No comments:

Post a Comment