Hi Catherine,

Have you shown the data to xtriage? (run by some pipelines at Diamond, or 
available as a standalone program in Phenix). The output might give you some 
hints.

You don't say what your current r free is at the moment, but if your phases are 
decent enough, I'd set modelcraft off on the data and go and get some lunch. 
That software is imo akin to witchcraft and has saved me a lot of manual 
tinkering to deliver a mostly complete asymmetric unit. It's found extra copies 
I didn't know I had, and even revealed proteolytic fragments that had been 
crystallised (I don't know how modelcraft behaved with coiled coils, bit 
probably still worth a punt).

I'd also run Zanuda to check the space group (I always do this now, having been 
tricked by crystals in the past).

Best of luck!

Dave


--

Dr David C. Briggs CSci MRSB (he/him)

Principal Laboratory Research Scientist, and

Co-chair, Crick Staff Consultative forum

Signalling and Structural Biology Lab

The Francis Crick Institute

London, UK

Working hours Mon-Fri 0900-1700

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"Would it not be better if one could really 'see' whether molecules...were just 
as experiments suggested?"
– Dorothy Hodgkin
________________________________
From: CCP4 bulletin board <[email protected]> on behalf of Catherine Back 
<[email protected]>
Sent: Friday, February 6, 2026 11:48:48 AM
To: [email protected] <[email protected]>
Subject: [ccp4bb] Crystal pathology diagnosis please


External Sender: Use caution.

Morning All,

My colleague and I are trying to solve the structure of a long, alpha helical 
bundle protein. We thought it had solved well with 2 molecules in the AU (which 
had decent stats), but upon a closer look there are very large 'solvent 
channels' in the lattice, with green/blue density within. It looks like there 
are protein molecules, positioned lengthways to the molecules in the AU, within 
these channels which the autoprocessing (at Diamond Light Source) is not 
picking up. See images attached.

I now have the image files and have been reprocessing them using xia2-DIALS in 
ccp4i2.

I am unsure what kind of crystal pathology we're dealing with here. There is no 
twinning warnings or anything else really that the processing has identified. 
The space group is always identified as P212121, unit cell 34.8, 184.8, 202.3, 
90.0, 90.0, 90.0.

I've tried quite a few different things to get the software to fit molecules 
into the density, including manually extending the unit cell, but all the 
outputs have been wrong or had really bad stats. I'm wondering if the indexing 
is wrong, but I'm not sure how to sort this. There is clearly something going 
on here, which is either really obvious (that I'm not identifying because I'm 
an idiot) or it's something more complex.

Any ideas/help will be much appreciated.

Best wishes,
Catherine Back


Dr Catherine R. Back (she/her)

Senior Post-doctoral Research Associate

School of Biochemistry

University of Bristol

UK


[email protected]


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