Purification, crystallization and preliminary X-ray diffraction analysis of the Kelch-like motif region of mouse Keap1.
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 P6(1) or P6(5), with unit-cell parameters a = b = 102.95, c = 55.21 A, and contains one molecule in the asymmetric unit. A complete diffraction data was collected to 2.25 A resolution using an R-AXIS IV++ imaging plate mounted on an RA-Micro7 Cu Kalpha rotating-anode X-ray generator.
Padmanabhan
Balasundaram
B
RIKEN Genomic Sciences Center, Tsurumi, Yokohama 230-0045, Japan.
Scharlock
Maria
M
Tong
Kit I
KI
Nakamura
Yoshihiro
Y
Kang
Moon-Il
MI
Kobayashi
Akira
A
Matsumoto
Takehisa
T
Tanaka
Akiko
A
Yamamoto
Masayuki
M
Yokoyama
Shigeyuki
S
D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D003460:Crystallization; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli; D000072019:Kelch-Like ECH-Associated Protein 1; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification; D014961:X-Ray Diffraction
D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D003598:Cytoskeletal Proteins; C117301:Keap1 protein, mouse; D000072019:Kelch-Like ECH-Associated Protein 1; D011993:Recombinant Fusion Proteins