It is possible that some Fe was turned to Ni isotope, which are very toxic
to those small animals. I learnt this story from a beamline scientist at
BNL. Not very sure, please validate it with your beamline scientist.

On Mon, Sep 8, 2025 at 9:07 AM Kevin Jin <[email protected]> wrote:

> Morning Catherine,
>
> Here was my explanation to my colleagues before, just a suggestion for
> your consideration.
>
> This could be a common case for the structures of a heme coordinating with
> Fe ion. Xtal structure is a thym-dynamic on average. In heme-Fe structures,
> it could be a mixture of Fe(II)/Fe(III). In some higher resolution
> structures(>1.23Ang), you can even tell the heme-plan bending like a
> pyramid with the Fe on the top of the bent heme plane, other than a
> symmetric octahedral geometry. A green cloud could always be observed in
> the Fe center.  It depends on the ratio of Fe(III).
>
> Based on a normal resolution of protein structures (1.5 - 2.5Ang), indeed
> a model,  it is difficult to define an exact bond length with its deviation
> at atomic level, like classic chemical crystallography. Therefore, what the
> heme structure showed is an up-down jumping between Fe(II) and Fe(III)
> with the Jahn–Teller effect (or the pseudo Jahn–Teller effect) .
>
> Because the respective positions of Fe(III)  and Fe(II) can not be exactly
> defined,  there may be a little portion Fe(III) in the heme structure. You
> may try to model the structure starting with (98% Fe(II)  + 2% Fe(III)) and
> refine the occp. Of course, the library  needs to be redefined for
> heme-Fe(III), but I won't suggest you do it. It is very time consuming.
>
> Good luck with your project.
>
> Kevin
>
> PS. you may put a bran new Iron nail in the protein solution. You may get
> a chance to reduce or eliminate the size of the green bubble.
>
>
>
>
>
>
>
>
>
>
> On Fri, Sep 5, 2025 at 2:40 AM Catherine Back <
> [email protected]> wrote:
>
>> Good morning,
>>
>> I am currently solving a structure containing four heme b molecules (res
>> 1.7 Å), each coordinated by two histidines. The refinement is looking good,
>> but the output from refinement has marked the Fe ions of each heme with
>> positive density (green) in the FoFc difference map - see image. Any ideas
>> why? I used the Monomer Library in Coot to add hemes ('HEM') in. Is it
>> something to do with the oxidation state of the Fe? And if so, is there
>> anything I can do about it?
>>
>> Best wishes,
>> Catherine
>>
>> Dr Catherine R. Back (she/her)
>>
>> Senior Post-doctoral Research Associate
>>
>> School of Biochemistry
>> University of Bristol
>>
>> UK
>>
>>
>>
>>
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