SPONTANEOUS ACHILLES EXPLOSIONS!
Proteomics to the rescue! (By the way, there is this whole series of bizarre children's books where there will be some silly problem and it's all COWS TO THE RESCUE or something. It's funny by the 11th page and continues through the 6th book somehow.)
Obviously, this group wants to understand why sometimes people's achilles up and explode just for fun, and they are able to get samples from patient who end up getting correction surgeries! Obviously, this is yet another place where genomics/transcriptomics of the tissue will probably tell you nothing - so it's proteomics time!
Interstingly the group breaks out iTRAQ 8-plex and does pooling, offline fractionation (by SCX, I think, but I forget now, did a singing daycare drop between reading it and now) and then analysis of the fractions on a Q Exactive (Classic, I think). All the files are up on PRIDE where they should be. I have no issue with iTRAQ 8-plex here, by the way. They turned up the collision energy, fragmented each target 2x before putting it on the dynamic exclusion list. The 8-plex allows them to run the QE at maximum (vendor permitted) speed with 17,500 resolution at m/z 200.
What I do have an issue with is the surprising pictures of the operation itself!! I was expecting a calibration or volcano plot and - blech.
Seriously, though, it is all pretty intersting. There are structural differences in the ruptured tissue that are clearly visible and they go into depth with IHC. They find a panel of targets that might be indicative of potential rupture candidates? It's a super compelling study all around - on something I now get to think about.
Interestingly...I think this is a dataset that would be a solid candidate for a reanalysis because it looks like this group didn't consider common collagen PTMs. I'm assuming when they considered dynamic oxidations they exclusively mean methionines. Collagens are hyper-modified. In fact, in the BOLT cloud search engine, Amol wrote in a whole crapload of common collagen PTMs in the first pass search because they're just that common. I think he got that idea from the cRAP database guy, whatever his name is. 😇
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