Hans,
Most natural toxins from snakes, scorpions etc are
50+/-some peptides. And quite a few of those have been
studied and crystallized (see pdb for a list). Having
worked on one of these structures as a graduate student, I
can share my experience:
- Purification is harder than you would think. You are
talking about < 10kD, usually around 5kD. Many methods
(size exclusion, even concentration over a simple
membrane) don't work as easily as you would like.
- I did not have much of a problem crystallizing (i.e. no
worse than other proteins, maybe even a little easier)
- Crystals tend to diffract well (maybe better than
average)
- Structures can be hard to solve; MIR is very difficult
because ions tend to not go into such crystals easily
(because the molecules are small and tightly packed?); MR
is hard because (again) it does not work very well on very
small systems
- Crystallization is not necessarily purification - if you
have a mixture of peptides to start with, it may be harder
to crystallize, or not: you might get a crystal that is a
(random-ish) mixture.
- If you have more than two cysteines in your sequence
(natural toxins typically do), the additional problem is
to get the correct folding and disulphide bridges;
alternatively it is very hard to discriminate between
correctly and incorrectly linked disulphides
Finally:
These sequence should be small enough for NMR. That may or
may not answer your questions, but it avoids your original
question.
Mark
-----Original Message-----
From: H. Raaijmakers
<[log in to unmask]>
To: CCP4BB
<[log in to unmask]>
Sent: Thu, Nov 10, 2011 8:16 am
Subject: [ccp4bb] crystallization of synthetic peptides
Dear crystallographers,
Because of the low cost and speed of synthesizing 40- to 60-mer peptides,
I wonder whether anyone has (good or bad) experiences crystalizing such
peptides. In literature, I've found up to 34-mer synthetic coiled coils,
but no other protein class. I can imagine that a protein sample with a few
percent "random deletion mutants" mixed into it won't crystallize easily,
but has anyone actually tried?
cheers,
Hans