Purification, crystallization and preliminary X-ray analysis of the galacto-N-biose-/lacto-N-biose I-binding protein (GL-BP) of the ABC transporter from Bifidobacterium longum JCM1217.
A recombinant galacto-N-biose-/lacto-N-biose I-binding protein (GL-BP) from Bifidobacterium longum JCM1217 has been prepared and crystallized by the hanging-drop vapour-diffusion method using 10 mg ml(-1) purified enzyme, 0.01 M zinc sulfate, 0.1 M MES buffer pH 5.9-6.4 and 20-22%(v/v) PEG MME 550 in the presence of 5 mM disaccharide ligands. Suitable crystals grew after 10 d incubation at 293 K. The crystals belong to space group C222(1), with unit-cell parameters a = 106.3, b = 143.6, c = 114.6 A for the lacto-N-biose I complex and a = 106.4, b = 143.4, c = 115.5 A for the galacto-N-biose complex, and diffracted to 1.85 and 1.99 A resolution, respectively.
Wada
Jun
J
Graduate School of Biostudies, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan.
Suzuki
Ryuichiro
R
Fushinobu
Shinya
S
Kitaoka
Motomitsu
M
Wakagi
Takayoshi
T
Shoun
Hirofumi
H
Ashida
Hisashi
H
Kumagai
Hidehiko
H
Katayama
Takane
T
Yamamoto
Kenji
K
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D001644:Bifidobacterium / Q000737:chemistry*; D003460:Crystallization; D017931:DNA Primers; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D014961:X-Ray Diffraction
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D017931:DNA Primers; D011994:Recombinant Proteins