Visualization of conformational transition of GRP94 in solution

Author:

Sun Shangwu1ORCID,Zhu Rui1,Zhu Mengyao1ORCID,Wang Qi1ORCID,Li Na2ORCID,Yang Bei134ORCID

Affiliation:

1. Shanghai Institute for Advanced Immunochemical Studies and School of Life Science and Technology, ShanghaiTech University

2. National Facility for Protein Science in Shanghai, Shanghai Advanced Research Institute (Zhangjiang Laboratory), Chinese Academy of Sciences, Shanghai, China

3. Shanghai Frontiers Science Center for Biomacromolecules and Precision Medicine, ShanghaiTech University

4. Shanghai Clinical Research and Trial Center, Shanghai, China

Abstract

GRP94, an ER paralog of the heat-shock protein 90 family, binds and hydrolyses ATP to chaperone the folding and maturation of its selected clients. Compared with other hsp90 proteins, the in-solution conformational dynamics of GRP94 along the ATP hydrolysis cycle are less understood, hindering our understanding of its chaperoning mechanism. Leveraging small-angle X-ray scattering, negative-staining EM, and hydrogen–deuterium exchange coupled mass-spec, here we show that in its apo form, ∼60% of mouse GRP94 (mGRP94) populates an “extended” conformation, whereas the rest exist in either “close V” or “twist V” like “compact” conformations. Different from other hsp90 proteins, the presence of AMPPNP only impacts the relative abundance of the two compact conformations, rather than shifting the equilibrium between the “extended” and “compact” conformations of mGRP94. HDX-MS study of apo, AMPPNP-bound, and ADP-bound mGRP94 suggests a conformational transition from “twist V” to “close V” upon ATP binding and a back transition from “close V” to “twist V” upon ATP hydrolysis. These results illustrate the dissimilarities of GRP94 in conformation transition during ATP hydrolysis from other hsp90 paralogs.

Funder

MOST | National Natural Science Foundation of China

Science and Technology Commission of Shanghai Municipality

Publisher

Life Science Alliance, LLC

Subject

Health, Toxicology and Mutagenesis,Plant Science,Biochemistry, Genetics and Molecular Biology (miscellaneous),Ecology

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