Tuesday, November 19, 2019
A wild TMTPro Paper has appeared!!
About darned time!
Just accepted at JPR -- the first (as far as I'm aware, please correct me if I'm wrong, study showing the use of TMTPro (previously TMT16-plex)!
Quick summary of my rapid readthrough:
1) This group typically uses NCE of 38 for TMT10/11-plex reagents, they use 32 for TMTPro (please keep in mind that proper HCD NCE can vary from system to system and there are ways to calibrate for that now. The important part is that the HCD is lower/closer to what we use for unlabled peptides! This is particularly good for those of us still using MS2 for TMT. The authors describe the use of both MS2 and SPS MS3 on their instrument. (And -- in my hands an HCD of 32 on an Orbitrap Fusion lines up pretty close to a NCE HCD of 27 on a Q Exactives -- again, varies from instrument to instrument, but this all sounds right to me!)
2) The larger tag makes the peptides a bit more hydrophobic (elute later) but it is a shift of a few minutes that can be easily adjusted for
3) When comparing number of peptide/protein IDs directly TMTPro results in a few percent less identifications, but you get 5 extra samples done simultaneously, so I still call that a win.
Concise, well-executed little study that will deserve the thousands of citations it will get for being the first one to press.
And -- for those of us dying to get our hands on a TMTPro dataset -- all these files have been deposited! (PRIDE PXD014750) I'm filling out the form on PRIDE now to have the files released for public download.