Also check if the mean value of the map over the ASU is zero. If so, the map 
was probably calculated without the 0,0,0 term of the Fourier series, so zero 
does not represent zero density but rather the mean density.

On 03/04/2019 12:43 PM, Ian Tickle wrote:

Hi Sen

If you multiply the electron density in a voxel by the voxel volume you should 
get an estimate of the number of electrons contained in that voxel, and then 
you can add up the numbers of electrons in all the voxels occupied by an atom 
to get the total number of electrons in that atom, which is basically the same 
as what you are saying.

However note that the electron density is a point sample: it's not the average 
density in the voxel, so the above calculation won't be quite accurate, 
depending on the 'smoothness' of the density.  This is like the error in an 
integral by use of Simpson's rule.  To be sure of accounting for all the 
density you need to sample it finely, say not more that Dmin/4.

There are two more issues here: first the atomic positions are never error-free 
which reduces the contribution to the density by the factor D in the expression 
2mFo-DFc (or mFo for centric phases) for the map coefficients.  So if the 
errors were sufficiently large D would tend to zero and you would get no 
density at all!  The density that you see is really only that part of the true 
density for which there is evidence in the experimental data.

Second, what exactly do you mean by "add all voxel densities around an atom"?  
The electron density could easily extend 2 Ang. from an atomic centre, depending on the 
atom's finite size (represented by the form factor), its thermal motion (B factor) and 
series termination effects (resolution).  So if you don't go out far enough you will fail 
to account for some fraction of the electron count.  The problem is of course you can't 
go so far as to overlap bonded atoms which will be well within 2 Ang. distance.  The 
standard method of dealing with this is to represent 'soft' atoms (where the distance 
between atoms may be less than the sum of their radii) as Voronoi polyhedra (like the 
packing of soap bubbles!).  Is that how you handled it?

Cheers

-- Ian


On Mon, 4 Mar 2019 at 15:47, Yao, Sen <[email protected] 
<mailto:[email protected]>> wrote:

    Hi all,

    I have been using the electron density maps available on the PDBe website 
to run some analysis. And I run into this question that I hope that I can get 
some help from the community.

    In the ccp4 format, the electron density is represented as a 3-d array map, 
with each number corresponds to the density value of a voxel in real space. If 
I add all voxel densities around an atom together and divided it by the number 
of electrons of that atom, in theory it should give me a ratio with the unit 
Å-3 (angstrom to the power of -3), and this ratio should be inversely related 
to the voxel volume. (Correct me if I am wrong here.) However, after I got this 
ratio for each atom, aggregated it into chains and calculated a median, and 
then compared the chain median to the voxel volume over all PDB structures with 
electron density available, they show a slope of ~1/3 instead of expected 1 
(see attached link). That is, almost all the values in the electron density 
maps are only about 1/3 of represented electrons.

        https://drive.google.com/file/d/1K_3uxZfUPuTdH1DtKJgKHLRUA5NHTVPB/view?usp=sharing 
<https://urldefense.proofpoint.com/v2/url?u=https-3A__drive.google.com_file_d_1K-5F3uxZfUPuTdH1DtKJgKHLRUA5NHTVPB_view-3Fusp-3Dsharing&d=DwMFaQ&c=ogn2iPkgF7TkVSicOVBfKg&r=cFgyH4s-peZ6Pfyh0zB379rxK2XG5oHu7VblrALfYPA&m=DQk9GIsPtYWv1IJTpGpK8ZW9vGk3XjlPB0GbBPxwYjo&s=z5pf9EEAoR1DrakQMp9nTAQ-Z4Sp5tigP5O4-Phm4WU&e=>


    So my question is, is there a conversion or scaling factor that PDBe uses 
to generate the ccp4 files? If so, is that information stored in the ccp4 files 
or anywhere else? And if not, why do I observe this 1/3 ratio pretty 
consistently across the whole PDB?

    I would really appreciate any insights on this matter. Thank you!

    Sincerely,
    Sen

    --

    Sen Yao, PhD

    Center for Environmental and Systems Biochemistry
    Markey Cancer Center
    University of Kentucky, Lexington KY


    
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