Dear All
checking out the Lysozyme crystallization methods on the web I liked the
Rigaku Instructions that I found:
(http://www.rigaku.com/protein/crystallization.html)
...create a drop of 3ul lysozyme solution, and 3 ul of well solution,
respectfully, for a total drop size of 6ul...
So
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Hi Martyn,
this recipe tends to work for me...
Lysozyme: 50 mg/ml in 0.1 M Sodium Acetate pH 4.8
Reagent: 8% w/v Sodium Chloride, 0.1 M Sodium Acetate pH 4.8
Additional Reagents: Index Reagent 8, 22, 28, 31, 34, 40, 58, 59, 69, 86, 88
Mix equal amounts of lysozyme with reagent, incubate at 4 or
Dear Martin,
The original procedure with polyethylene glycol comes from:
Stura, E.A. (1998) Strategy 3: Reverse Screening. In Crystallization of
Proteins: Techniques, Strategies and Tips. A laboratory manual (Bergfors,
T., ed) International University Line. pp. 113-124.
If you follow the
I was recently testing out the 30% PEGMME 5k, 0.1M NaOAc pH 4.5, 1M
NaCl method mentioned in a Hampton Research catalog and attributed to
Enrico Stura. I see that he has also just commented on this thread.
I found that at 80mg/ml, batch with 1.5ul:1.5ul protein to precipitant
ratio,
- I think the original poster was only calling attention to the fact
that some proteins want to be treated respectfully in order to
crystallise (and the fact that Rigaku Japan realises this). I find
that indeed the case.
Other proteins, however, prefer the attitude I don't know why I am
I was going to comment that I have learned the following: respect does not
mean the same thing in all places in the world. Some time back I had a protein
here that I thought needed extra respect and I had learned from a Rigaku
employee how to do this - I bowed very very deeply in front of the
I'd like to thank you all, the problem is resolved. Apparently, in the
pdb file, the carbonyl oxygen should be listed right after the carbonyl
carbon not after the side chain atoms! M
**
Mohd A. Salameh, Ph.D.
Mayo Clinic Cancer Center
Griffin Cancer
If respect is concerned, try to stop a blow of a sharp samurai sword with a
sharp word.
I myself have no problems whatsoever with the Rigaku recipes. And yes, I have
stopped,
at leas once, a blow of samurai sword with my bare hands. Since than I paint
pictures with my left foot.
BTW
In ALL my
It makes slight sense if the word 'respect' can be correlated with whether
the drop is mixed (protein+ppt). i have heard some protein may indeed
crystallize if it were not mixed or vice-versa (do not expect it with
lysozyme though). it would make sense if diffusion played a role but with
few ul
Hello,
Is there any way to search in AutoMR mode but have the
translation only being brute forced?
Regards,
F.
Sorry, I forgot to mention my previous message concerns using Phaser.
Francois Berenger wrote:
Hello,
Is there any way to search in AutoMR mode but have the
translation only being brute forced?
Regards,
F.
Steve,
My general strategy is to start with an ideal glycan (an Asn linked to
NAG-NAG-(MAN)3 ) and superimpose the Asn on the residue from my protein.
Then you can move the whole glycan as a rigid body until the Asn and
first NAG are roughly positioned. Then you can tweak any sugars further
Maybe is a mistake by a non-English speaker. Instead of respectfully should
say
respectively
Dear Steve,
I would also use Damian's approach, but the sequence of the core should be
NAG-NAG-BMA-(MAN)2. This is improtant because the correct stereochemistry
restraints for beta-mannose can only be applied when you call the residue
BMA.
Building carbohydrates also comes with special validation
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