Lisa,
As others have said, using careful data collection and the modern program
suites available (SHARP, Phenix, etc.), a 300 KD complex with 111 Se-Met
residues should be quite solvable. But you didn't state is what is in the
asymmetric unit (the important figure): one complex with 111 Se-Me
Hi Lisa, hi all,
Please do not discard the alternative method(s) of "conventional" heavy
atoms. Co-crystallisation or heavy-atom containing mother liquor soaks.
You may remember that "monster" complexes have been solved in the past
by such methods, and sometimes there are difficulties in cryst
On Tue, Jun 12, 2012 at 8:53 PM, Frank von Delft
wrote:
> Finding 111 sites should be feasible without other tricks than very careful
> data collection (see below); if you have two or more copies in the ASU, you
> may find you need to do what the ribosome guys did, namely use other
> derivatives
If you don't have crystals yet, you'll find getting it to crystallize is
your main problem.
Finding 111 sites should be feasible without other tricks than very
careful data collection (see below); if you have two or more copies in
the ASU, you may find you need to do what the ribosome guys di
If you want to use se-Met, you might want to start by labeling only one protein
at a time. For example, if you have A,B,C,D, grow crystals like this:
se-A, B, C, D
A, se-B, C, D,
etc.
Then try combinations of 2, then 3, then if you haven't got the phases you
need, try all 4.
And remember, if
Do you have crystals?
Do they diffract? If so, to what resolution?
What resolution do you require to answer your biological question?
F
On Jun 12, 2012, at 7:46 PM, LISA wrote:
> Hi all,
>
> My work is to solve huge complex containing 4 different proteins and total
> molecular weight is ab
Hi all,
My work is to solve huge complex containing 4 different proteins and total
molecular weight is about 300 KD. I can purify the complex by co-expression
them in E.coli. This complex contains 8 protein A, 2 protein B and 1
protein C and D. protein B and protein C have homology structures
de