Do you expect more than one molecule in the asymmetric unit?
Determined from the Matthews Coefficient (poor), size exclusion column (better), or self RF (best) ?
On Nov 7, 2013, at 8:36 AM, Zhihong Yu <[log in to unmask]> wrote:
> Hi, all
>
> I'm a rookie in resolving a brand new structure. I have some questions for my current case and look forward to some suggestions.
>
> Now I’m working on a protein like this: N-ter(55aa)—domainA(110aa)—linker(30aa)—domainB(150aa)—C-ter(20aa), I got a diffraction data just to 3.5Å, and there is no complete homology structure in pdb bank, but only a homology structure (named as structureX later) for domainB with ~30% sequence identity, so I have some questions as following:
>
> 1. Is it possible to find a resolution through MR approach using structureX as a search model? Especially considering that the resolution is only 3.5Å. Currently I just tried once using phaser and refine the structure, I can get a R/Rfree of 0.45/0.55, and it looks like most of backbone in the structureX, especially those within helix or sheet, can be well described by 2Fo-Fc density. Is this primary result promising or not?
>
> 2. If it’s possible, what’s the general optimal procedure I should follow?
>
> Really thanks for any advice and suggestions!
>
> Zhihong
>
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Francis E. Reyes PhD
215 UCB
University of Colorado at Boulder
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