An improved fluorescent tag and its nanobodies for membrane protein expression, stability assay, and purification

Hongmin Cai; Hebang Yao; Tingting Li; Cedric A. J. Hutter; Yanfang Li; Yannan Tang; Markus A. Seeger; Dianfan Li

doi:10.1038/s42003-020-01478-z

Communications Biology (Dec 2020)

An improved fluorescent tag and its nanobodies for membrane protein expression, stability assay, and purification

Hongmin Cai,
Hebang Yao,
Tingting Li,
Cedric A. J. Hutter,
Yanfang Li,
Yannan Tang,
Markus A. Seeger,
Dianfan Li

Affiliations

Hongmin Cai: University of Chinese Academy of Sciences, National Center for Protein Science Shanghai, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences
Hebang Yao: University of Chinese Academy of Sciences, National Center for Protein Science Shanghai, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences
Tingting Li: University of Chinese Academy of Sciences, National Center for Protein Science Shanghai, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences
Cedric A. J. Hutter: Institute of Medical Microbiology, University of Zurich
Yanfang Li: University of Chinese Academy of Sciences, National Center for Protein Science Shanghai, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences
Yannan Tang: University of Chinese Academy of Sciences, National Center for Protein Science Shanghai, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences
Markus A. Seeger: Institute of Medical Microbiology, University of Zurich
Dianfan Li: University of Chinese Academy of Sciences, National Center for Protein Science Shanghai, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences

DOI: https://doi.org/10.1038/s42003-020-01478-z
Journal volume & issue: Vol. 3, no. 1
pp. 1 – 16

Abstract

Read online

In this work, the authors demonstrate that a coral thermostable GFP (TGP) is an improved tag compared to conventional GFPs. In addition to reporting melting point stability at temperatures near 90 °C, its fusion also helps increase expression levels of test membrane proteins. They further generated synthetic nanobodies against TGP to facilitate purification.

Published in Communications Biology

ISSN: 2399-3642 (Online)
Publisher: Nature Portfolio
Country of publisher: United Kingdom
LCC subjects: Science: Biology (General)
Website: https://www.nature.com/commsbio/

About the journal