Reading that text I was convinced that this lady is indeed no intellect, just emotions. No facts or math, just gossip. And then she just concluded all women must be like that.

Nice work. Thanks for a fantastic skeetorial, always appreciated

I’ll let you know if we see anything interesting!

One thing is for sure, fun with RAF never ends! (10/10)

One thing took us by surprise: It seems that the asymmetric dimer comes at the end of the RAF activation cycle. We got more structures and saw that the Receiver looks the most like a canonical active kinase. Is the asymmetric conformation then the final catalytically active state of RAF? (9/10)

If we titrate those mutants into cells with wild type RAFs in the background, we should see a dose dependent effect. And we did. The beautiful model must be correct! (8/10)

But what is the difference between active asymmetric and active symmetric structures? We made BRAF mutants
(i) AAAA mutant = Receiver but not Activator
(ii) Kinase-dead mutant = Activator but not Receiver
(iii) Double mutant = neither
(7/10)

This work started by Yasushi Kondo solving an X-ray crystal structure of BRAF in complex with its substrate MEK1 where the BRAF dimeric interface was asymmetric, with the NtA motif of only one of the subunits making interface contacts. Yes, like in that beautiful model! (6/10)

However, to paraphrase the late Cyrus Chothia, if the data don't fit a beautiful model, you need more data. You can't just forget about the NtA - cancer genetics and the early Marais experiments don't allow it! (5/10)

The problem? Multiple BRAF and CRAF structures were solved since, and … dimers were symmetric, kinases looked active, and the NtA looked disordered. (4/10)

15 years later, Susan Taylor et al proposed a model: Active RAFs are dimers and one subunit (the "Activator") uses its NtA to activate the other subunit (the "Receiver"). It is only the Receiver that takes the fully active kinase conformation. tinyurl.com/RAFmodel (3/10)

In 1999, Marais lab showed that this is because of the 4-residue N-terminal acidic (NtA) motif. In CRAF, the NtA must be phosphorylated to become acidic and the kinase to become active. In BRAF the NtA is already acidic, making BRAF one step closer to (over)active. tinyurl.com/NtAmotif (2/10)

It’s the chicken or the egg question. We are only now realising this layer is important because it was so hard to study. And because of that, people studying it probably didn’t do well in their careers

😅