Working with very large or multi-domain proteins can feel like wrestling a spaghetti monster – they’re floppy, prone to aggregation and cellular degradation and thus often low in yield.
A classic structural biology trick for taming such divas is “divide and conquer.” Rather than expressing a hundreds of kDa beast in one piece, you might break the gene into domains or functional modules and express those separately. Truncated constructs can ensure higher stability during expression which dramatically improves protein yield. On top of that, removing unstructured regions or internal flexibilities often gets you a more tractable, crystallizable protein.
And as a bonus, well-behaved protein fragments often crystallize when the full-length protein would not, yielding valuable structural insights.
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