Selection of an RNA molecule that specifically inhibits the protease activity of subtilisin

Hiroshi Takeno, Seiji Yamamoto, Terumichi Tanaka, Yoshiyuki Sakano, Yo Kikuchi*

*この研究の対応する著者

研究成果: Article査読

12 被引用数 (Scopus)

抄録

RNA ligands (RNA aptamers) to a protease subtilisin were selected from pools of random RNA by SELEX (systematic evolution of ligands by exponential enrichment) and by use of a subtilisin-immobilized Sepharose column. After eight rounds of selection, RNA aptamers were isolated by cloning to a plasmid vector. We characterized one of the selected RNA molecules. This RNA aptamer displayed specific inhibition toward the subtilisin activity, even when the assay for subtilisin was performed using the chromogenic small peptide as substrate, and almost no inhibitory activity toward trypsin and chymotrypsin, although these enzymes are serine proteases similar to subtilisin. These findings indicate that this RNA can differentially recognize the surfaces of similar proteases. Kinetic analysis of the RNA aptamer revealed that the inhibition constant (K(i)) toward subtilisin was 2.5 μM.

本文言語English
ページ(範囲)1115-1119
ページ数5
ジャーナルJournal of Biochemistry
125
6
出版ステータスPublished - 1999
外部発表はい

ASJC Scopus subject areas

  • 生化学

フィンガープリント

「Selection of an RNA molecule that specifically inhibits the protease activity of subtilisin」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル