Open AccessPurification, crystallization and preliminary X‐ray diffraction analysis of the Kelch‐like motif region of mouse Keap1
Author(s) -
Padmanabhan Balasundaram,
Scharlock Maria,
Tong Kit I.,
Nakamura Yoshihiro,
Kang MoonIl,
Kobayashi Akira,
Matsumoto Takehisa,
Tanaka Akiko,
Yamamoto Masayuki,
Yokoyama Shigeyuki
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309104032506
Subject(s) - crystallization , motif (music) , x ray , crystallography , diffraction , x ray crystallography , materials science , chemistry , physics , optics , organic chemistry , acoustics
Keap1 (Kelch‐like ECH‐associating protein 1) is a negative regulator of the Nrf2 transcription factor in the cytoplasm. The Kelch/DGR (double‐glycine repeat) domain of Keap1 associates with Nrf2 as well as with actin filaments. A recombinant protein containing both the Kelch/DGR domain and the C‐terminal region of mouse Keap1 was expressed in Escherichia coli , purified to near‐homogeneity and crystallized by the sitting‐drop vapour‐diffusion method. The crystal belongs to space group P 6 1 or P 6 5 , with unit‐cell parameters a = b = 102.95, c = 55.21 Å, and contains one molecule in the asymmetric unit. A complete diffraction data was collected to 2.25 Å resolution using an R‐AXIS IV ++ imaging plate mounted on an RA‐Micro7 Cu K α rotating‐anode X‐ray generator.