Electron microscopic study of the human THOC1-THOC2-THOC3 subcomplex
10.3969/j.issn.1674-8115.2020.01.001
- Author:
Yong-Mei LIU
1
Author Information
1. Shanghai Institute of Precision Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine
- Publication Type:Journal Article
- Keywords:
MRNA nuclear transport;
Negative-staining electron microscopy;
THO complex;
TREX complex
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2020;40(1):1-7
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the structure of the human THOC1-THOC2-THOC3 subcomplex by negative-staining electron microscopy. Methods:The recombinant protein components were co-expressed in cultured insect cells, and the protein complex was isolated via sequential Ni-NTA and Strep-Tactin affinity purification followed by Superose gel filtration chromatography. The purified protein sample was then subjected to negative-staining electron microscopy and single particle image analysis. Results:The recombinant human THOC1-THOC2-THOC3 complex with high purity and good homogeneity was obtained by using a tandem affinity purification scheme with Ni-NTA and streptavidin-based chromatography. Preliminary study on the structure of human THOC1-THOC2-THOC3 subcomplex was achieved by electron microscopy using negative staining with uranium formate. Conclusion:A low resolution model of human THOC1-THOC2-THOC3 subcomplex was achieved by single particle reconstruction.
