High-performance affinity beads for identifying drug receptors.
We have developed a method using novel latex beads for rapid identification of drug receptors using affinity purification. Composed of a glycidylmethacrylate (GMA) and styrene copolymer core with a GMA polymer surface, the beads minimize nonspecific protein binding and maximize purification efficiency. We demonstrated their performance by efficiently purifying FK506-binding protein using FK506-conjugated beads, and found that the amount of material needed was significantly reduced compared with previous methods. Using the latex beads, we identified a redox-related factor, Ref-1, as a target protein of an anti-NF-kappaB drug, E3330, demonstrating the existence of a new class of receptors of anti-NF-kappaB drugs. Our results suggest that the latex beads could provide a tool for the identification and analysis of drug receptors and should therefore be useful in drug development.
Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
N
N
Shimizu
K
K
Sugimoto
J
J
Tang
T
T
Nishi
I
I
Sato
M
M
Hiramoto
S
S
Aizawa
M
M
Hatakeyama
R
R
Ohba
H
H
Hatori
T
T
Yoshikawa
F
F
Suzuki
A
A
Oomori
H
H
Tanaka
H
H
Kawaguchi
H
H
Watanabe
H
H
Handa
D000595:Amino Acid Sequence; D001483:Base Sequence; D016227:Benzoquinones / Q000378:metabolism*; D017931:DNA Primers; D006801:Humans; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D011422:Propionates / Q000378:metabolism*; D011955:Receptors, Drug / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D016227:Benzoquinones; D017931:DNA Primers; D016328:NF-kappa B; D011422:Propionates; D011955:Receptors, Drug; D011994:Recombinant Proteins; C075569:E 3330