Discover and read the best of Twitter Threads about #ubiquitin
Most recents (3)
In my 1st tweetorial, I’m happy to share our preprint—my first, first author manuscript! We used @10xGenomics single-nucleus RNA-seq in a mouse model of electroconvulsive seizures (ECS) to assess activity-regulated gene expression in hippocampal #neurons. biorxiv.org/content/10.110…
Activity-regulated gene expression patterns in the #hippocampus regulate #plasticity, #learning, and #memory, and are linked to risk and treatment response for neuropsychiatric disorders. However, the cell type-specificity of these patterns is still not well characterized.
We sequenced a bunch of neuronal nuclei (15,990) from mouse hippocampus and used graph-based clustering and a priori marker genes to annotate these nuclei across all major hippocampal subregions and neuron types in both of our experimental conditions (Sham and ECS). 
As promised, a 🧵 to convince you to consider SP4 for your next #proteomics sample prep!
TL;DR: Cell lysate -> pure peptides ready for LC-MS with 20-30 min handling time w/o any filters (FASP, iST, S-Trap) or specialist beads (SP3). 1/n
biorxiv.org/content/10.110… @biorxivpreprint
TL;DR: Cell lysate -> pure peptides ready for LC-MS with 20-30 min handling time w/o any filters (FASP, iST, S-Trap) or specialist beads (SP3). 1/n
biorxiv.org/content/10.110… @biorxivpreprint
Starting off a lab (babraham.ac.uk/our-research/s…) where proteomics was going to be a major approach, our first recruit—postdoc @HarveyEJohnston—set off to craft our work-horse sample prep work-flow.
2/n
2/n
This workflow needed to be: (1) quick-ish; (2) cheap-ish; (3) versatile & scalable (e.g., different sample amounts & vols); (4) able to clean up detergents; (5) compatible with #PTM enrichment approaches (especially #ubiquitin TUBEs & diGly); (3) easy to teach to students.
3/n
3/n
Incredible paper from the Elledge group in @CellCellPress, cell.com/cell/fulltext/…, discovering new regulation of protein stability by “DesCEND” degrons and shaping of eukaryotic proteome... 1/18
Starts with proteome-scale discovery of linear peptide degrons (sequences mediating degradation) - peptide fusion with GFP, co-expressed with DsRED as internal control - sort based on GFP/DsRED ratio, low ratio = unstable GFP = peptide acting as degron... 2/18
CRL (cullin ring E3 ligase) inhibition stabilized a fraction of peptide-GFP fusions - shown to be mostly Cul2 and Cul5 dependent. CRISPR screening leads to some CRL2 adaptors... 3/18
