Functionalization of the BCL6 BTB domain into a noncovalent crystallization chaperone

• Main Authors: Thomas Zacharchenko, Stephanie Wright
• Format: Article
• Language: English
• Published: International Union of Crystallography 2021-03-01
• Series: IUCrJ
• Subjects: protein crystallization; crystallization chaperone; btb domain; porous crystal lattice
• Online Access: http://scripts.iucr.org/cgi-bin/paper?S2052252520015754

The production of diffraction-quality protein crystals is challenging and often requires bespoke, time-consuming and expensive strategies. A system has been developed in which the BCL6 BTB domain acts as a crystallization chaperone and promiscuous assembly block that may form the basis for affinity-capture crystallography. The protein of interest is expressed with a C-terminal tag that interacts with the BTB domain, and co-crystallization leads to its incorporation within a BTB-domain lattice. This strategy was used to solve the structure of the SH3 domain of human nebulin, a structure previously solved by NMR, at 1.56 Å resolution. This approach is simple and effective, requiring only routine protein complexation and crystallization screening, and should be applicable to a range of proteins.