<div dir="ltr">Recreating the profile was just a sanity check. Really I want to look at conservation across the protein of interest.</div><br><div class="gmail_quote"><div dir="ltr" class="gmail_attr">On Thu, 15 Feb 2024 at 16:13, Peter Cock <<a href="mailto:p.j.a.cock@googlemail.com">p.j.a.cock@googlemail.com</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left:1px solid rgb(204,204,204);padding-left:1ex">You may have more joy starting from the HMM for the model?<br>
<br>
<a href="https://www.ebi.ac.uk/interpro/wwwapi//entry/pfam/PF08241?annotation=hmm" rel="noreferrer" target="_blank">https://www.ebi.ac.uk/interpro/wwwapi//entry/pfam/PF08241?annotation=hmm</a><br>
<br>
Peter<br>
<br>
On Thu, Feb 15, 2024 at 4:06 PM Dan Bolser <<a href="mailto:dan.bolser@outsee.co.uk" target="_blank">dan.bolser@outsee.co.uk</a>> wrote:<br>
><br>
> Searching HMMER gives me (cryptic?) details of the model match:<br>
><br>
> Family-Id Family-Accession Clan Start End Ali-Start Ali-End Model-Start Model-End Bit-Score Ind.-E-value Cond.-E-value Description<br>
> ==================================================================================================================================<br>
><br>
> Methyltransf_11 PF08241.15 CL0063 63 173 63 172 1 95 51.30 1.5e-13 3.1e-17 Methyltransferase domain<br>
><br>
> ---------------------------------------------------------------------------------------------------------------------------------<br>
><br>
> MODEL LdvGcGtGrlaealakrg.arvvgvDlskemlklakekaseeglkvefvvadaeklpfednsfDlvvssevlhhv...e............dpekalkeiaRvLkpgGllv<br>
> MATCH L +GcG+ +l+ +l g +v+ vD+s ++++ +++ + ++++ +d++kl f+++sfD+v+ + +l+ + e ++++l+e+ RvL pgG+++<br>
> PPL 569************7779*************77766666666.69*****************************8662667778888888******************97<br>
> SEQ LVLGCGNSALSYELFLGGfPNVTSVDYSSVVVAAMQARHAHVP-QLRWETMDVRKLDFPSASFDVVLEKGTLDALlagErdpwtvssegvhTVDQVLSEVSRVLVPGGRFI<br>
><br>
><br>
> So I guess it's possible to work backwards from there...<br>
><br>
><br>
><br>
> On Thu, 15 Feb 2024 at 16:02, Peter Cock <<a href="mailto:p.j.a.cock@googlemail.com" target="_blank">p.j.a.cock@googlemail.com</a>> wrote:<br>
>><br>
>> Sorry yes, I used this as a threshold for if a column was gappy, and<br>
>> likely to be in the model or not:<br>
>><br>
>> amino_acids.get("-", 0) < len(align)*0.5<br>
>><br>
>> Might need to use <= to match, or perhaps they use a more<br>
>> sophisticated criteria.<br>
>><br>
>> Peter<br>
>><br>
>> On Thu, Feb 15, 2024 at 3:56 PM Dan Bolser <<a href="mailto:dan.bolser@outsee.co.uk" target="_blank">dan.bolser@outsee.co.uk</a>> wrote:<br>
>> ><br>
>> > Sorry, by 'half gaps' I thought you meant a specific thing, like, "look at that half gap!". I guess you mean, columns where half of the values in the column are gaps.<br>
>> ><br>
>> > Looking directly at the alignment, I guess there is no way to work out precisely which columns are gaps or not in the model?<br>
>> ><br>
>> ><br>
>> ><br>
>> ><br>
>> ><br>
>> ><br>
>> > On Thu, 15 Feb 2024 at 15:47, Dan Bolser <<a href="mailto:dan.bolser@outsee.co.uk" target="_blank">dan.bolser@outsee.co.uk</a>> wrote:<br>
>> >><br>
>> >> Hi Peter,<br>
>> >><br>
>> >> On Thu, 15 Feb 2024 at 15:28, Peter Cock <<a href="mailto:p.j.a.cock@googlemail.com" target="_blank">p.j.a.cock@googlemail.com</a>> wrote:<br>
>> >>><br>
>> >>> Hello Dan,<br>
>> >>><br>
>> >>> It that is a probability at the end, you have the wrong denominator -<br>
>> >>> should be len(align) == sum(amino_acids.values()<br>
>> >><br>
>> >><br>
>> >> Makes no difference, they are the same (including -s). The profile reports -s as 'Insert Probability'.<br>
>> >><br>
>> >><br>
>> >>> Note this seed alignment has 164 columns, yet the model page says the<br>
>> >>> model has 95 columns. If I count the number of columns which are half<br>
>> >>> gaps that's pretty close...<br>
>> >><br>
>> >><br>
>> >> I don't understand what that means or implies... Is something wrong somewhere?<br>
>> >><br>
>> >><br>
>> >> Cheers,<br>
>> >> Dan.<br>
>> >><br>
>> >><br>
>> >>> Peter<br>
>> >>><br>
>> >>> On Thu, Feb 15, 2024 at 3:09 PM Dan Bolser <<a href="mailto:dan.bolser@outsee.co.uk" target="_blank">dan.bolser@outsee.co.uk</a>> wrote:<br>
>> >>> ><br>
>> >>> > Hi,<br>
>> >>> ><br>
>> >>> > Sorry, I can't follow the docs (or find the right docs).<br>
>> >>> ><br>
>> >>> > I've got the 'seed' stockholm alignment for this domain:<br>
>> >>> > <a href="https://www.ebi.ac.uk/interpro/entry/pfam/PF08241/entry_alignments/?type=seed" rel="noreferrer" target="_blank">https://www.ebi.ac.uk/interpro/entry/pfam/PF08241/entry_alignments/?type=seed</a><br>
>> >>> ><br>
>> >>> > and I'm trying to reproduce the signature it shows here:<br>
>> >>> > <a href="https://www.ebi.ac.uk/interpro/entry/pfam/PF08241/logo/" rel="noreferrer" target="_blank">https://www.ebi.ac.uk/interpro/entry/pfam/PF08241/logo/</a><br>
>> >>> ><br>
>> >>> > I'm not sure a) why the probabilities differ in the profile relative to the seed alignment, or b) how to filter columns in the alignment by those that have a match in the model (see columns 4-6 in the alignment, which are gaps in the model).<br>
>> >>> ><br>
>> >>> > I think if I can answer b) then the answer to a) will be, "look at the full alignment".<br>
>> >>> ><br>
>> >>> > Here is my crude 'best guess' code:<br>
>> >>> ><br>
>> >>> > import gzip<br>
>> >>> > import Bio.AlignIO<br>
>> >>> ><br>
>> >>> > # msa = "PF08241.alignment.full.gz"<br>
>> >>> > msa = "PF08241.alignment.seed.gz"<br>
>> >>> ><br>
>> >>> > with gzip.open(msa, "rt") as handle:<br>
>> >>> > align = Bio.AlignIO.read(handle, "stockholm")<br>
>> >>> > ncols = align.get_alignment_length()<br>
>> >>> ><br>
>> >>> > for col in range(ncols):<br>
>> >>> > amino_acids = dict()<br>
>> >>> > for s in align[:, col]:<br>
>> >>> > amino_acids[s] = amino_acids.get(s, 0) + 1<br>
>> >>> > print(amino_acids)<br>
>> >>> > for s in amino_acids:<br>
>> >>> > print(f"{s}: {amino_acids[s]:3d} {amino_acids[s] / len(align):.3f}")<br>
>> >>> ><br>
>> >>> ><br>
>> >>> ><br>
>> >>> > I have the feeling I'm doin it rong...<br>
>> >>> ><br>
>> >>> > The above is just a 'warm up', really I want to see the conservation score, base by base on a given protein in the alignment (where it matches the model).<br>
>> >>> ><br>
>> >>> > Many thanks for any suggestions, and sorry for not being able to find the right document to answer these questions.<br>
>> >>> ><br>
>> >>> ><br>
>> >>> > kthxbi,<br>
>> >>> > Dan.<br>
>> >>> > _______________________________________________<br>
>> >>> > Biopython mailing list - <a href="mailto:Biopython@biopython.org" target="_blank">Biopython@biopython.org</a><br>
>> >>> > <a href="https://mailman.open-bio.org/mailman/listinfo/biopython" rel="noreferrer" target="_blank">https://mailman.open-bio.org/mailman/listinfo/biopython</a><br>
</blockquote></div>