Hi folks

I can’t help feeling that if there are data extending to 0.97Å and there are 
multiple conformations/multiple components occupying the same region of (real) 
space/non-unity occupancies, the problem is crying out for SHELXL…

Harry

> On 27 Apr 2020, at 07:53, Schreuder, Herman /DE <[email protected]> 
> wrote:
> 
> Dear Abhishek,
> 
> I did not follow the links given by Paul. However the way I proceeded in 
> these cases was to first generate an alternative conformation for the problem 
> residues, save the file and then do the mutation and save the mutated file. 
> Then, using an editor, I would cut the alternative conformation from the 
> original residue and paste in the alternate conformation (e.g. conformation 
> B) from mutated residue.
> 
> It is not very elegant, but it works (if coot and refmac cooperate).
> 
> Best, Herman
> 
> -----Ursprüngliche Nachricht-----
> Von: CCP4 bulletin board <[email protected]> Im Auftrag von Abhishek Anan
> Gesendet: Sonntag, 26. April 2020 21:42
> An: [email protected]
> Betreff: [EXTERNAL] Re: [ccp4bb] modelling MET (methionine) and SME 
> (met-sulfoxide)
> 
> EXTERNAL : Real sender is  [email protected]   
> 
> 
> 
> Dear all,
> 
> Thanks for the suggestions. It is synthetic peptide so the residue identity 
> is unambiguous.
> 
> I am not clear on how to model both MET and SME in coot, do a real space 
> refinement and then save the file for refinement in phenix. I tried alternate 
> conformation and then mutating one of them but that didn't work as both 
> conformations were mutated. What I also just noticed is that the refinement 
> of structure with just SME results in positive densities around all side 
> chain carbons even with the SME.cif loaded into phenix. What could be wrong 
> here?
> 
> best wishes,
> Abhishek
> 
> 
> 
> 
> 
> On 4/26/20, Paul Emsley <[email protected]> wrote:
>> 
>> On 26/04/2020 16:21, Abhishek Anan wrote:
>>> Dear all,
>>> 
>>> I have a peptide crystal structure at 0.97 Å that contains two 
>>> surface exposed Methionine. The CE atoms of both MET have a 
>>> suspiciously high b-factor >40 and a positive density. In addition, 
>>> the sulfur atom SD has a large negative density (b-factor ~23).
>>> 
>>> I initially suspected that the MET may have oxidized to MET-sulfoxide 
>>> and tried to model only the MET-sulfoxide. This again resulted in 
>>> negative density.
>>> 
>>> I think that the peptides might be partly oxidized which brings me to 
>>> my question. Is there a way to model both MET and MET-sulfoxide into 
>>> the density much like alternate conformation with options to refine 
>>> their respective occupancies.
>> 
>> 
>> Yes. This is called micro-heterogeneity
>> 
>> And is documented here:
>> 
>> https://urldefense.proofpoint.com/v2/url?u=https-3A__www.wwpdb.org_doc
>> umentation_procedure&d=DwIFaQ&c=Dbf9zoswcQ-CRvvI7VX5j3HvibIuT3ZiarcKl5
>> qtMPo&r=HK-CY_tL8CLLA93vdywyu3qI70R4H8oHzZyRHMQu1AQ&m=CYQUM_mrfiCDADJ1
>> onMNLQYYLwXD23pcMOTm7KnoNkM&s=A_ke05wSRD1nm9vQxFwLPCzpmUAWRTVlaVkVSTRw
>> z8M&e=
>> 
>> That should "just work" if you then give the model to refmac.
>> 
>> FWIW, Coot is, AFAIR, not 100% happy with such models.
>> 
>> Paul.
>> 
>> 
>> 
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