Enhanced protein secretion from insect cells by co-expression of the chaperone calreticulin and translation initiation factor eIF4E.

Molecular Biotechnology
Chao-Yi TengTzong-Yuan Wu

Abstract

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.

References

Oct 6, 1999·Current Opinion in Biotechnology·T A Kost, J P Condreay
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
Oct 9, 2002·EMBO Reports·Gia K VoeltzTom A Rapoport
Feb 15, 2003·Biochimica Et Biophysica Acta·Christopher G TateD Clive Williams
Mar 4, 2003·Nature Reviews. Molecular Cell Biology·Lars Ellgaard, Ari Helenius
May 3, 2003·Biochemical and Biophysical Research Communications·Rai KneeMarek Michalak
May 13, 2003·The Journal of Biological Chemistry·Liyan ZhangGunilla Olivecrona
Feb 26, 2004·Journal of Immunological Methods·Jean-Marie FourneauPeter M van Endert
Oct 19, 2004·Journal of Virological Methods·Stephen A KabaMonique M van Oers
Apr 5, 2005·Methods : a Companion to Methods in Enzymology·Marie-Eve PaquetDavid B Williams
Jun 25, 2005·Current Opinion in Cell Biology·Christine M FederovitchRandolph Y Hampton
Aug 9, 2005·Biochemical and Biophysical Research Communications·Ying-Ju ChenTzong-Yuan Wu
Sep 1, 2005·Biotechnology Letters·Eun-Young YunO-Yu Kwon
Jun 30, 2006·Current Opinion in Cell Biology·Nica BorgeseErik Snapp
Jul 7, 2006·Annual Review of Cell and Developmental Biology·Sebastián BernalesPeter Walter
Jun 15, 2007·Nature Reviews. Molecular Cell Biology·David Ron, Peter Walter
Apr 26, 2008·Plant & Cell Physiology·Anna ChristensenMarianne Sommarin
Jul 15, 2009·Cell Cycle·Peter M Fischer
May 6, 2010·Biotechnology and Applied Biochemistry·Richard B HitchmanLinda A King
Dec 14, 2011·Trends in Biochemical Sciences·Christoph BieniossekImre Berger

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