In situ structure and dynamics of an alphacoronavirus spike protein by cryo-ET and cryo-EM
Journal
Nature Communications
Journal Volume
13
Journal Issue
1
Date Issued
2022-12-01
Author(s)
Huang, Cheng Yu
Draczkowski, Piotr
Wang, Yong Sheng
Chang, Chia Yu
Chien, Yu Chun
Cheng, Yun Han
Wu, Yi Min
Wang, Chun Hsiung
Chang, Yuan Chih
Yang, Tzu Jing
Tsai, Yu Xi
Khoo, Kay Hooi
Hsu, Shang Te Danny
Abstract
Porcine epidemic diarrhea (PED) is a highly contagious swine disease caused by porcine epidemic diarrhea virus (PEDV). PED causes enteric disorders with an exceptionally high fatality in neonates, bringing substantial economic losses in the pork industry. The trimeric spike (S) glycoprotein of PEDV is responsible for virus-host recognition, membrane fusion, and is the main target for vaccine development and antigenic analysis. The atomic structures of the recombinant PEDV S proteins of two different strains have been reported, but they reveal distinct N-terminal domain 0 (D0) architectures that may correspond to different functional states. The existence of the D0 is a unique feature of alphacoronavirus. Here we combined cryo-electron tomography (cryo-ET) and cryo-electron microscopy (cryo-EM) to demonstrate in situ the asynchronous S protein D0 motions on intact viral particles of a highly virulent PEDV Pintung 52 strain. We further determined the cryo-EM structure of the recombinant S protein derived from a porcine cell line, which revealed additional domain motions likely associated with receptor binding. By integrating mass spectrometry and cryo-EM, we delineated the complex compositions and spatial distribution of the PEDV S protein N-glycans, and demonstrated the functional role of a key N-glycan in modulating the D0 conformation.
SDGs
Type
journal article