Hi

It depends on the software you’re using for the alignment. Some does a “better” 
job than others (e.g. I’d expect a strictly sequence based alignment like Blast 
to do a “good” job, but I’ve seen occasional misalignments [that are obvious to 
the eye!] with some of the HMM-based software like HHAlign - I have no idea why 
this might happen because the group that produces HH-suite doesn’t appear to be 
active enough to address issues on their github; I know that many of the 
authors are no longer working on the project). 

So, my follow-up question would be “which software are you using for this?”.

Harry

> On 11 Feb 2026, at 05:27, Thripthi Shenoy 
> <[email protected]> wrote:
> 
> Greetings to all,
> 
> I am currently working on understanding the structure-based sequence 
> alignment of the crystal structure of NOXA BH3 domain and the complete 
> protein structure obtained from AlphaFold. According to my understanding, the 
> BH3 binding domain from the crystal structure should get aligned with the BH3 
> domain in the entire protein structure. However, this is not the case and the 
> identical amino acids of the BH3 domains are not getting aligned. I would be 
> grateful to know if this is a correct approach to proceed with the structure 
> based protein alignment.
> 
> Thanking you in advance
> With regards,
> Thripthi S.
> 
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