🏖️Back from an amazing vacation in 🌴Sanya🐟. Refreshed and recharged, ready to dive back to research!

🤣 I found this fun picture from a few years back. To ensure high coverage of the protein truncation library, each plate can have 50K - 100K colonies 🦠. 🧵5/6

🛡️ I also mapped how molecular chaperones (cellular "helpers") guide folding at specific sites. Trigger factor and DnaK showed varying effects along the full-length protein. This opens new doors for understanding protein misfolding diseases. 🧵4/6

🔍 Surprise finding: Homologous protein domains (EF-G vs EF-Tu) fold through completely different pathways! EF-Tu forms a stable intermediate halfway through synthesis, while EF-G waits until the very end. Structural topology matters! 🧵3/6

⚡ My approach combines SecM arrest peptide, dual fluorescent reporters, cell sorting, and custom deep sequencing to capture folding "snapshots" across entire protein sequences. ⚛ Think of it as a molecular camera that catches the exact moment a protein finds its shape! 🧵2/6

🤔 How do proteins fold while they're still being made? As the main project in my PhD, I developed "Arrest Peptide Profiling" to record proteins folding with as they emerge from ribosomes in living cells 🦠 - with single codon resolution! @natcomms.nature.com‬ 🧵 1/6
#ProteinFolding

Over the moon at #SMPS4 in Bolzano.

Photographer @jritmejeris.bsky.social

Interestingly, the permutation of PTMs on the two possible tyrosine residues generated drastically different signals, enabling highly accurate localization of the PTMs.

Because of the charge difference between tyrosine sulfation (-1) and phosphorylation (-2), we were able to accurately identify the PTMs on the PSK peptide, even when the two variants were highly similar in mass.

We conjugated the plant peptide hormone PSK with a strand of DNA on its N-terminus and placed 15 aspartate on its C-terminus. While the helicase pulls the molecule through the nanopore, we observed dramatic current increase due to the high local salt induced by dense charges.

Very happy to share our latest work @acsnano
on applying #nanopore to protein analysis!
With the motor protein Hel308, we translocate DNA-peptide conjugate through MspA and successfully distinguish sulfation and phosphorylation at #singlemolecule level.