RNA aptamers that reversibly bind photoresponsive azobenzene-containing peptides.
Modulation of biological networks assembled by diverse interactions among biologically active molecules has provided a platform for innovative biotechnologies. Here, we report RNA aptamers that bind to a photoresponsive peptide (KRAzR; Lys-Arg-azobenzene-Arg) containing azobenzene chromophore, which can change its structure by photoirradiation. Aptamers were identified after 10 cycles of an in vitro selection procedure starting with a DNA library containing a 70 nt random region. Surface plasmon resonance (SPR) analysis demonstrated that interactions between aptamers and KRAzR were fully controlled by appropriate photoirradiation to the SPR sensor chip. Upon irradiation of 360 nm on the KRAzR-immobilized surface, the binding of each aptamer to the surface was significantly decreased. Subsequent photoirradiation of the same surface with 430 nm restored the aptamer binding to the surface. We also observed that direct photoirradiation of the aptamer-peptide complex on a gold surface actively promoted dissociation of the complex. Furthermore, a doped reselection method was applied to acquire structural and sequence information of aptamer 66. From a data analysis of the conserved region and the mutation frequency, we were able to select a plausible secondary structure among three candidates predicted by computational folding simulation.
Hayashi
Gosuke
G
Department of Regulatory Bioorganic Chemistry, The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki 567-0047, Japan.
Hagihara
Masaki
M
Nakatani
Kazuhiko
K
D000595:Amino Acid Sequence; D052157:Aptamers, Nucleotide / Q000235:genetics / Q000378:metabolism*; D001391:Azo Compounds / Q000737:chemistry*; D001483:Base Sequence; D006046:Gold / Q000737:chemistry; D007536:Isomerism; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism*; D055668:Photochemical Processes; D013499:Surface Properties
D052157:Aptamers, Nucleotide; D001391:Azo Compounds; D009842:Oligopeptides; D006046:Gold; C009850:azobenzene