Open AccessCrystallization and preliminary X‐ray crystallographic studies of the coiled‐coil domain of PIST
Author(s) -
Shin YoungCheul,
Seo Eun Kyoung,
Jeon JuHong,
Park Hyun Ho
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113007082
Subject(s) - coiled coil , crystallization , crystallography , pdz domain , domain (mathematical analysis) , diffraction , fusion protein , chemistry , c terminus , escherichia coli , resolution (logic) , x ray crystallography , physics , materials science , biochemistry , amino acid , recombinant dna , optics , computer science , organic chemistry , gene , mathematical analysis , mathematics , artificial intelligence
PIST [PDZ (PSD‐95, Discs‐large and ZO‐1) protein interacting specifically with TC10] functions as a regulator of membrane trafficking with Rab6A. Recently, the involvement of the fusion of PIST with ROS1 in cancer development has been identified. In this study, the coiled‐coil domain of PIST, which is the domain responsible for interaction with Rab6A and fusion with ROS1, corresponding to amino acids 29–133, was overexpressed in Escherichia coli using engineered C‐terminal His tags. The coiled‐coil domain of PIST was then purified to homogeneity and crystallized at 293 K. Finally, X‐ray diffraction data were collected to a resolution of 4.0 Å from a crystal belonging to the hexagonal space group P 6 2 22 or P 6 4 22, with unit‐cell parameters a = b = 85.19, c = 240.09 Å, γ = 120.00°.