Wednesday, December 17, 2014
Proteomic assessment of wound healing
Whew...finally stopped for lunch and got away from that PC monitor long enough to get on my tablet (sigh...) and read a really cool paper!
I knew this one would be great when I saw how many words in the title of the article that I probably couldn't define if asked right on the spot. The paper is called "In Vivo Assessment of Protease Dynamics in Cutaneous Wound Healing by Degradomics Analysis of Porcine Wound Exudates" and comes from Fabio Sabino et al.,
The pivotal words here? Degradomics and exudates. This study actually uses a pig wound healing model and uses a really interesting (and complex) quantitative technique to determine the level of protease activity in these tissues. I skipped the methods section regarding how they get the pig wounds. I'm not that much of biologist.
The sample prep technique is a combination of iTRAQ 4-plex and TAILS (terminal amine isotopically labeling of substrates. You can read about TAILS at this Wikipedia page here (and I think somewhere on this silly blog as well). This figure pretty much sums it up, though.
Doing TAILS gives you an idea of the level of proteolysis in your different samples. But this group stepped it up a level by throwing in iTRAQ 4-plex as well, so we can get quan on this analysis and plex more samples. Its worth noting that they didn't invent this technique (called iTRAQ-TAILS), these guys did in 2011, but its a new one to me.
All the MS analysis was done with the always awesome (and aging gracefully) workhorse, the Orbitrap XL. This is a really cool study. If having a really interesting biological model and impressively complex experimental design wasn't enough, they really do a nice job of their post processing analysis. The pathways and heatmaps are concise and well-constructed. All around, this is just a really nice paper.
You can grab it here, where it is currently open access at MCP.