Photo-cross-linked small-molecule microarrays as chemical genomic tools for dissecting protein-ligand interactions.
We have developed a unique photo-cross-linking approach for immobilizing a variety of small molecules in a functional-group-independent manner. Our approach depends on the reactivity of the carbene species generated from trifluoromethylaryldiazirine upon UV irradiation. It was demonstrated in model experiments that the photogenerated carbenes were able to react with every small molecule tested, and they produced multiple conjugates in most cases. It was also found in on-array immobilization experiments that various small molecules were immobilized, and the immobilized small molecules retained their ability to interact with their binding proteins. With this approach, photo-cross-linked microarrays of about 2000 natural products and drugs were constructed. This photo-cross-linked microarray format was found to be useful not merely for ligand screening but also to study the structure-activity relationship, that is, the relationship between the structural motif (or pharmacophore) found in small molecules and its binding affinity toward a protein, by taking advantage of the nonselective nature of the photo-cross-linking process.
Kanoh
Naoki
N
Antibiotics Laboratory, Discovery Research Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. hisyo@riken.jp
Asami
Aya
A
Kawatani
Makoto
M
Honda
Kaori
K
Kumashiro
Saori
S
Takayama
Hiroshi
H
Simizu
Siro
S
Amemiya
Tomoyuki
T
Kondoh
Yasumitsu
Y
Hatakeyama
Satoru
S
Tsuganezawa
Keiko
K
Utata
Rei
R
Tanaka
Akiko
A
Yokoyama
Shigeyuki
S
Tashiro
Hideo
H
Osada
Hiroyuki
H
D001667:Binding, Competitive; D003432:Cross-Linking Reagents / Q000737:chemistry*; D023281:Genomics / Q000379:methods*; D005898:Glass; D006838:Hydrocarbons / Q000737:chemistry; D008024:Ligands; D008697:Methane / Q000031:analogs & derivatives* / Q000737:chemistry; D019920:Photoaffinity Labels / Q000737:chemistry; D010777:Photochemistry; D040081:Protein Array Analysis; D011506:Proteins / Q000737:chemistry*; D011869:Radioligand Assay; D013329:Structure-Activity Relationship; D013499:Surface Properties; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D006838:Hydrocarbons; D008024:Ligands; D019920:Photoaffinity Labels; D011506:Proteins; C030011:carbene; D008697:Methane