Institute of Genetics and Developmental Biology, Cas

Mingjiang Chen

Bing Wang

Guifu Liu

Jiayang Li

Xiangbing Meng

Hong Yu

Yanhui Jing

Huihui Liu

The disclosure discloses a method for preparing a multisubunit SCF E3 ligase with a fusion protein through in vitro reconstitution, and a use of the multisubunit SCF E3 ligase. The method for preparing a multisubunit SCF E3 ligase with a fusion protein through in vitro reconstitution disclosed by the disclosure includes: subjecting an engineered SKP1-Cullin1-RBX1 fusion protein (referred to as eSCR) to a reaction with an F-box protein in a reaction system to obtain the multisubunit SCF E3 ligase, where the eSCR fusion protein has an amino acid sequence from position 10 to position 1062 of SEQ ID NO: 2. Experimental results show that different multisubunit SCF E3 ligases are successfully prepared with the eSCR fusion protein through in vitro reconstitution in the disclosure; the reconstituted multisubunit E3 ligase has biological activity.

Method for preparing multisubunit SCF E3 ligase with fusion protein through in vitro reconstitution, and use of multisubunit SCF E3 ligase

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Beijing, China

Mingjiang Chen