Wednesday, August 22, 2018

Chemical crosslinking in vivo in plants!!!


Somebody seriously just asked me about this recently and I came up with nothing. And here it is!!

Plant proteomics sucks. Give it a run sometime!

Annnnd....Another peptide from...{{drumrolllll}}.....RUBISCO!
(I can't find the originator of this Rubisco gif, just random repostings for karma. Happy to credit appropriately)

Okay -- so how on earth would you use chemical crosslinking to do interactomics? You stop thinking about it and just do it this way.



Would I have thought to introduce a crosslinker under vacuum? Nope! 

Would I have used inexpensive and powerful dimethyl labeling? Nope! (I only just found out what a nice reagent this is. Another scientist in my group generates beautiful data with it) 

How do you get rid of the infuriating rubisco? 

Extensive fractionation
Gradient centrifugation
And enrichment (...cause you use a crosslinker that can be enriched!)

It takes a lot for me to post a paper about that Scumbag Arabidopsis, but this is useful enough that it could definitely be applied to real plants and things that matter! 

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