Re: [phenixbb] Autosol: MIRAS

This one is solvable, but with extreme difficulty. I recently completed a structure solution with experimental phases starting at 5.0 A using phase information from multiple derivatives. How would you describe a somewhat satisfactory map? F On Sep 5, 2012, at 7:08 PM, [email protected] wrote:

Hi Shya,

I did wavelength scan, got a good signal for Se and used appropriate wavelengths for data collection and also used experimental f' and f'' values for phasing. I think the reasons SAD or MAD for SeMet data is not working are (i) low resolution: 3.7 for SeMet (Anomalous data is up to 4.8A) (ii) I should have mentioned this earlier: 3 out of 6 Se are very close to N-terminus, possible they are disordered. Unit cell is also some what big..100, 120 and 320A; F222 space group.

AK

-----Original Message----- From: Shya Biswas To: PHENIX user mailing list Sent: Thu, Sep 6, 2012 7:29 am Subject: Re: [phenixbb] Autosol: MIRAS

Hi AK, Did you do a wavelength scan when you collected the SE dataset you need to put the values of f' and f'' from your wavelength scan in order to locate the heavy atom sites, 6 methionine should be enough to phase your molecule. Shya

On Wed, Sep 5, 2012 at 9:25 PM, wrote:

...

Hi all,

I am trying to solve a structure through experimental phasing using AUTOSOL. I have a couple of heavy atom derivative datasets (Hg, La, Eu, Cd) and also a SeMet data. Unfortunately all the datasets are of low resolution (3.7-4.2A) and there are possibly 4-8 molecules in the asu. MIR, SAD and MAD alone did not give any convincing solution.

However, MIRAS, with a combination of few heavy atom datasets and the anomalous data from SeMet crystals, gave a somewhat satisfactory map. But the heavy atom site picked by AUTOSOL list only one of the heavy atoms i.e. Lanthanum. In another set of run, the solution of which was not convincing, the heavy atom substructure had only Hg. There are 6 Met out of 200 residues in one molecule and mass spec results show that Se incorporation is 100%.

Now, my doubt is that why does the heavy atom substructure contain only La and how can I get the substructure involving Se from this solution (or the datasets used)? Se location is going to help me a lot for finding a starting point to assign side chains.

Any suggestion would be greatly appreciated.

Thanks AK

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