Expression and membrane integration of SARS-CoV E protein and its interaction with M protein |
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Authors: | Shih-Chi Chen Shih-Yen Lo Hsin-Chieh Ma Hui-Chun Li |
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Affiliation: | (1) Graduate Institute of Molecular and Cellular Biology, Tzu Chi University, Hualien, Taiwan;(2) Graduate Institute of Medical Biotechnology, Tzu Chi University, Hualien, Taiwan;(3) Graduate Institute of Medical Sciences, Tzu Chi University, 701, Section 3, Chung Yang Road, Hualien, Taiwan |
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Abstract: | The severe acute respiratory syndrome (SARS)-CoV E gene fragment was cloned and expressed as a recombinant protein fused with a myc tag at the N-terminus in vitro and in Vero E6 cells. Similar to other N-glycosylated proteins, the glycosylation of SARS-CoV E protein occurred co-translationally in the presence of microsomes. The SARS-CoV E protein is predicted to be a double-spanning membrane protein lacking a conventional signal peptide. Both of the transmembrane regions (a.a. 11–33 and 37–59) are predicted to be α-helices, which penetrate into membranes by themselves. As expected, these two transmembrane regions inserted a cytoplasmic protein into the endoplasmic reticulum membrane. Either of these two transmembrane domains co-localized with M protein. Both the transmembrane domains of E protein are required to interact with M protein, while either of the hydrophilic regions (a.a. 1–10 or 60–76) is dispensable as shown by co-immunoprecipitation assay. These results are important for the study of SARS-CoV assembly. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | SARS-CoV E protein Co-translational Membrane integration Protein– protein interactions |
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