<font color='black' size='2' face='arial'>@<font size="2" color="black" face="arial">Gabor: Thanks Gabor! I'll try MR-SAD. Hope it works. <br>
<br>
@Andre: I have tried Shelx and Phenix. Se sites from both of them were not the same. I tried giving Shelxd output to Phenix also. That also did not help. I have not tried </font><font size="2" color="black" face="arial">Autosharp </font><font size="2" color="black" face="arial">yet. I'll see that too..<br>
<br>
However, what now I am primarily looking at is the way to get Se-substructure from the MIRAS run. Is there a way to get that? autosol gives only one heavy atom in the output! Since I have a map which looks promising, so before going to the output of another run I want to explore all the possibilities from this. Se locations in this map should help. <br>
<br>
AK<br>
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<div style="font-family:arial,helvetica;font-size:10pt;color:black">-----Original Message-----<br>
From: Gabor Bunkoczi <gb360@cam.ac.uk><br>
To: PHENIX user mailing list <phenixbb@phenix-online.org><br>
Sent: Thu, Sep 6, 2012 5:52 am<br>
Subject: Re: [phenixbb] Autosol: MIRAS<br>
<br>
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<pre style="font-size: 9pt;"><tt>Dear AK,
if you just need the Se positions, you can run Phaser MR-SAD (available
from the GUI) using your partial model as a starting point, together with
the Se-Met data. Dependending on the model quality (and the model does not
have to be very good), it may be able to locate the Se-sites.
BW, Gabor
On Sep 6 2012, <a href="mailto:ash.k@aol.com">ash.k@aol.com</a> wrote:
> Satisfactory map means: I am expecting a coiled coil or helix bundle type
> of assembly and I could see some densities appropriate for helices. There
> are proper solvent channels and continuous stretches of densities.
>
> More to add about data on this: the data is anisotropic and the longer
> 'c' axis and alignment of helical density along c axis support this. This
> also makes me think that perhaps map is sensible.
>
> Few rounds of model building, refinement and DM has been successful to
> assign around 10 polyala helices and their distribution looks sensible
> from packing point of view. Now the problem is to assign the side chain.
> I was hoping to make use of Se locations for this. R and Rfree are still
> random, in the range of 50%. I am not sure, but it could be perhaps
> because most of the scattering material is still unassigned.
>
>
>
>
>
>
>-----Original Message-----
>From: Francis E Reyes <<a href="mailto:Francis.Reyes@colorado.edu">Francis.Reyes@colorado.edu</a>>
>To: PHENIX user mailing list <<a href="mailto:phenixbb@phenix-online.org">phenixbb@phenix-online.org</a>>
>Cc: phenixbb <<a href="mailto:phenixbb@phenix-online.org">phenixbb@phenix-online.org</a>>
>Sent: Thu, Sep 6, 2012 8:07 am
>Subject: 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, <a href="mailto:ash.k@aol.com">ash.k@aol.com</a> 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 <<a href="mailto:shyabiswas@gmail.com">shyabiswas@gmail.com</a>>
>To: PHENIX user mailing list <<a href="mailto:phenixbb@phenix-online.org">phenixbb@phenix-online.org</a>>
>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, <<a href="mailto:ash.k@aol.com">ash.k@aol.com</a>> 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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--
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Dr Gabor Bunkoczi
Cambridge Institute for Medical Research
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