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Combining cross‐crystal averaging and MRSAD to phase a 4354‐amino‐acid structure
Author(s) -
Busby Jason Nicholas,
Lott J. Shaun,
Panjikar Santosh
Publication year - 2016
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.374
H-Index - 138
ISSN - 2059-7983
DOI - 10.1107/s2059798315023566
Subject(s) - phaser , crystallography , resolution (logic) , crystal structure , multiplicity (mathematics) , chemistry , physics , mathematics , optics , computer science , geometry , artificial intelligence
The B and C proteins from the ABC toxin complex of Yersinia entomophaga form a large heterodimer that cleaves and encapsulates the C‐terminal toxin domain of the C protein. Determining the structure of the complex formed by B and the N‐terminal region of C was challenging owing to its large size, the non‐isomorphism of different crystals and their sensitivity to radiation damage. A native data set was collected to 2.5 Å resolution and a non‐isomorphous Ta 6 Br 12 ‐derivative data set was collected that showed strong anomalous signal at low resolution. The tantalum‐cluster sites could be found, but the anomalous signal did not extend to a high enough resolution to allow model building. Selenomethionine (SeMet)‐derivatized protein crystals were produced, but the high number (60) of SeMet sites and the sensitivity of the crystals to radiation damage made phasing using the SAD or MAD methods difficult. Multiple SeMet data sets were combined to provide 30‐fold multiplicity, and the low‐resolution phase information from the Ta 6 Br 12 data set was transferred to this combined data set by cross‐crystal averaging. This allowed the Se atoms to be located in an anomalous difference Fourier map; they were then used in Auto‐Rickshaw for multiple rounds of autobuilding and MRSAD.