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But structural biology can be very inefficient. Let's say you want to know which residues in a protein are important for a particular function, so you can twea
by DrScientist 6y ago
But structural biology can be very inefficient.
Let's say you want to know which residues in a protein are important for a particular function, so you can tweak them to improve function.
You could
- spend months or years solving a structure of the complex then look
at the structure and look at the direct contacts - then mutate to various other resides to
see if they affect function ( finding out that even with a structure of
the complex your chosen list is far from 100% correct )
- look at sequence alignments and then make a library of millions of variants and screen in a couple of months.
The first feels better as the glorious scientist is doing 'rational design', however it can be much more hit and miss and slower and more expensive.
As I understand it, one of the things about Cryo-EM is it's potentially much faster/easier than previous methods - but even so, looking at the structure doesn't get you to the answer direct - it's still very much hit and miss.
- natechols 6y agoBrute-forcing the problem with a variant library and high-throughput screening could easily end up being far more inefficient. At least with crystallography it's easy to set up the experiment, whether or not it actually works.
- dekhn 6y agoYou can even buy robots that do grid searches for crystallography (IE, if you're desperate, can't find conditions to make a crystal, and you've exhausted your postdocs, the robot will be able to try more possibilities in shorter time). Unfortunately they don't work very well and end up being a real pain to clean.
- entee 6y ago“Easy” is rarely a word I’ve heard associated with crystallography. Hence cryo-em! Easier and faster sample prep.
- natechols 6y agoIn the very best case, it is indeed easy! I've seen the entire process take less than a month for a completely novel structure - those were extremely lucky cases of course, but my point is that it costs very little to set up some screens and see what happens. (Small globular proteins of course, not ribosomes or ion channels.)
- entee 6y agoNice! I always assumed unless there was a known crystal, you’d be in for pain. “Good and globular” is the way to go I guess.