Enhanced protein secretion from insect cells by co-expression of the chaperone calreticulin and translation initiation factor eIF4E
| Autor: | Chao-Yi Teng, Tzong-Yuan Wu, Shou-Lin Chang, Monique M. van Oers |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
glycoprotein
Insecta ribosome entry site Recombinant Fusion Proteins Green Fluorescent Proteins Laboratory of Virology Golgi Apparatus Bioengineering calnexin Endoplasmic Reticulum Applied Microbiology and Biotechnology Biochemistry Laboratorium voor Virologie quality-control Genes Reporter Calnexin Animals Molecular Biology biology pathway Endoplasmic reticulum EIF4E endoplasmic-reticulum Alkaline Phosphatase PE&RC Fusion protein infection Cell biology Internal ribosome entry site Eukaryotic Initiation Factor-4E Secretory protein er messenger-rna Chaperone (protein) biology.protein Calreticulin Baculoviridae baculovirus expression system Molecular Chaperones Biotechnology |
| Zdroj: | Molecular Biotechnology, 54(1), 68-78 Molecular Biotechnology 54 (2013) 1 |
| ISSN: | 1073-6085 |
| Popis: | Host protein synthesis is shut down in the lytic baculovirus expression vector system (BEVS). This also affects host proteins involved in routing secretory proteins through the endoplasmic reticulum (ER)-Golgi system. It has been demonstrated that a secretory alkaline phosphatase–EGFP fusion protein (SEFP) can act as a traceable and sensitive secretory reporter protein in BEVS. In this study, a chaperone, calreticulin (CALR), and the translation initiation factor eIF4E were co-expressed with SEFP using a bicistronic baculovirus expression vector. We observed that the intracellular distribution of SEFP in cells co-expressing CALR was different from co-expressing eIF4E. The increased green fluorescence emitted by cells co-expressing CALR had a good correlation with the abundance of intracellular SEFP protein and an unconventional ER expansion. Cells co-expressing eIF4E, on the other hand, showed an increase in extracellular SEAP activity compared to the control. Utilization of these baculovirus expression constructs containing either eIF4E or CALR offers a significant advantage for producing secreted proteins for various biotechnological and therapeutic applications. |
| Databáze: | OpenAIRE |
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