In vitro selection of nonnatural ribozyme-catalyzing porphyrin metalation.
A new macromolecular catalyst, which is composed of nonnatural ribonucleotide, was synthesized by the in vitro selection (SELEX) method. A pool of RNAs consisting of random sequences was obtained by transcription from a pool of synthetic random-sequence DNAs in the presence of 2'-aminocytidine triphosphate (2'-amino-CTP) instead of CTP. The pool was incubated with N-methylmesoporphyrin-immobilized gel; bound nonnatural RNAs were then collected and amplified by reverse-transcription following the polymerase chain reaction. The amplified DNAs were again transcribed in the presence of 2'-amino-CTP and applied to the gel. This selection process was repeated 10 times. The selected RNAs were cloned and sequenced. The RNAs not only bound to the ligand, N-methylmesoporphyrin but also catalyzed the metalation reaction of porphyrin.
Kawazoe
N
N
Department of Biological Science and Technology, Faculty of Engineering, University of Tokushima, Tokushima 770-8506, Japan.
Teramoto
N
N
Ichinari
H
H
Imanishi
Y
Y
Ito
Y
Y
D001483:Base Sequence; D002384:Catalysis; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008670:Metals / Q000737:chemistry; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011166:Porphyrins / Q000737:chemistry / Q000378:metabolism*; D016337:RNA, Catalytic / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012641:Selection, Genetic; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D046911:Macromolecular Substances; D008670:Metals; D011166:Porphyrins; D016337:RNA, Catalytic