RNA aptamers that specifically bind to the Ras‐binding domain of Raf‐1

FEBS Letters

Shirouzu, Mikako¹  Hirao, Ichiro²  Yokoyama, Shigeyuki³  Sakamoto, Kensaku³  Kimoto, Michiko³ 

[1] Cellular Signaling Laboratory, The Institute of Physical and Chemical Research (RIKEN), Hirosawa, Wako-shi, Saitama 351-0198, Japan;Yokoyama CytoLogic Project, ERATO, JST, c/o RIKEN, Hirosawa, Wako-shi, Saitama 351-0198, Japan;Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan

关键词: Raf-1;    Ras;    RNA aptamer;    In vitro selection;    ERK;    extracellular signal-regulated kinase;    FPLC;    fast protein liquid chromatography;    GST;    glutathione S-transferase;    MAPK;    mitogen-activated protein kinase;    RBD;    Ras-binding domain;    MEK;    MAPK/ERK kinase;    GTPγS;    guanosine 5′-O-(3-thiotriphosphate);   

DOI  :  10.1016/S0014-5793(98)01572-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

RNA aptamers that bind to the Ras-binding domain (RBD) of a proto-oncogene product, Raf-1, were isolated from a pool of random sequences using a glutathione S-transferase-fused RBD (GST-RBD). The RNA molecules bind to the GST-RBD, but not to GST, with dissociation constants of about 300 nM. In contrast, these RNA aptamers do not bind to the Ras-binding domain of the RGL protein, which is also known to be activated by Ras. The aptamers actually compete with Ras for binding to the Raf-1 RBD. The anti-Raf-1 aptamers may be used to specifically inhibit the Ras-Raf interaction in the complicated signaling network in mammalian cells.

【 授权许可】

Unknown