It’s interesting that the 8600 doesn’t appear to be marketed with single cell capabilities compared to other vendors. I wonder why SCIEX didn’t at least have a note in this space?

This sounds fantastic and can’t wait to try it on some new peptidomics data!

Congratulations Atul and team! Amazing body of work.

Yes probably redundant, but there are considerations before pushing back. Do you have lysate remaining or would you have to repeat experiments? How many other ‘battles’ are there in the comments? If there are bigger fights and you have lysate than might be worth a ‘quick’ WB to appease the gods.

Thanks for all the support @unimelbcmr.bsky.social!!

Big thanks to all our collaborators for making this wok possible. @unimelbcmr.bsky.social @craigagoodman1.bsky.social and many others…

Why do we care? Because we have previously shown that UFC1 levels are negatively correlated with lean mass in various mouse strains, and knockdown of UFC1/UFMylation can increase muscle function. Maybe myosin UFMylation is a key link? elifesciences.org/articles/82951

We quantified UFMylation in the above human biopsies revealing several increased UFMylation sites including Myosins! Additional mapping of sites on Myosins reveal they are extensively modified, and in silico modelling suggest sites adjacent to ATP binding plays a functional role.

What did we find?... A whole bunch of cool substrates including UFMylation of Myosins which we validate following knockdown of UFC1 (the E2 ligase) in vivo combined with TMT-16plex labelling and LC-MS/MS…

In collaboration with CST, we developed antibodies that recognise "remnant ValGly" left on UFM1-conjugates following tryptic digestion. Similar to the remnant ubiquitin "diGly" approach published by www.cell.com/molecular-ce...

However, check out this anti-UFM1 blot from control muscle biopsies and people with ALS... A big band at 30kDa (probably RPL26) but clearly more to discover including big heavy bands >250kDa!

UFMylation is a ubiquitin like modification of UFM1 applied to substrate lysine residues. <15 substrates have been identified (a nice summary: www.cell.com/molecular-ce.... RPL26 has been most extensively characterised and suggested to be the major substrate…

Very nice indeed. In this example does LLOQ=LLOD?

The grant agencies in Australia seem to specifically announce outcomes on Friday afternoons. I think the idea is so you can cool off over the weekend. However, better than Monday morning!

Yes, this is where things are interesting. All databases produce similar number of total peptides. Large db’s identify more novel peptides missing in canonical db but the larger search space and requirement for higher score at 1%FDR means peptides get filtered out.

Yes, I think you are right. The different databases have different levels of redundancy. I’ll search some variant databases to confirm. Thank you!

Congratulations and nice collection of spirits 👌