Congratulations!! 🥳

Thanks Reid! Looks like they are queued to release with the next PDB update on Jan. 15

Thanks Stephanie! Hope all is going will w/your new lab - excited to see all the amazing work to come!

Thanks Joe!

Thanks, glad you enjoyed it!

Thanks Alex!

Tagging all the folks who I can find here... @amanglik.bsky.social @willowcoyote.bsky.social @justingenglish.bsky.social @delemottelab.bsky.social ...

There is certainly more than can be shared here, but I will end it there with a HUGE thanks to everyone involved! This was both a challenging and fun project to be involved in with many great folks without which none of this would be possible. (13/13)

Ultimately, this enabled us to identify residues which change pronation with conformation and pH. Combining our DMS, cryo-EM, MD, and extensive pharmacologic validation (by XP Huang), we were able to devise a comprehensive model for pH activation of GPR68 (12/n)

Well, Darko went above and beyond. He developed a new method which uses coevolutionary information to identify both alternative receptor conformations AND the effect of pH on the conformational ensemble. (11/n)

We needed an inactive-state structure model to fully interpret our results, so we teamed up with
Darko Mitrovic (in @delemottelab.bsky.social lab) to do some molecular dynamics simulations of GPR68 (10/n)

We followed up with a surface expression screen to determine the which mutations alter receptor expression. This allowed to deconvolve the mutational effects on expression vs activation to uncover GOF and LOF activity specific to activation. (9/n)

The result: a comprehensive map of each mutation's effect on pH activation. (8/n)

We then generated a mutational library of GPR68 using the DIMPLE platform developed in @willowcoyote.bsky.social's lab and screened it using our cAMP FACS assay (7/n)

Using GPR68 as our model, we set out to use mutational scanning to determine the effect of every mutation on proton activation. We developed a new FACS-based method to measure Gs coupled receptor activation (enabled by a new TRE's from @justingenglish.bsky.social ) (6/n)

We needed to ascribe a functional role for each residue in proton activation. Unfortunately, these receptors are littered with an enormous number polar and charged residues which may be implicated. (5/n)