Two topics today...
1) Problem with LIBCHECK.
Thanks to all who replied to my issue. I was able to resolve the issue
before the flurry of responses came in. Basically, I took my CSA modified
amino acid, aligned it as best I could to the CYS (especially at the CA),
and then manually edited the PDB
Greetings fellow Crystallographers,
I'm working on a structure at 1.8-A resolution that contains an acetone
crosslink between 2 cysteines (crosslink was incorporated by adding
1,3-dichloroacetone). I figured that the easiest way to model this is to
mutate one of the cysteines to S-acetonylcysteine
Refold protein 8M Urea:
Sanjiv,
The recommendations for rapid dilution and trying 6M GdmCl are great
recommendations. As an alternative, I had a similar problem years ago with a
membrane-associated receptor domain that was mostly found in inclusion
bodies after expression (I assume this is your
This post has a question and an answer (good karma)...answer first
In response to Matt's question about periplasmic harvesting, we do an
osmotic lysis of e.coli by a series of centrifugations. First, harvest your
cells normally (4k 15 min). Then vigorously resuspend in a 20% (w/v)
sucrose, 1 mM ED
