Enhanced protein expression in the baculovirus/insect cell system using engineered SUMO fusions.

Protein Expression and Purification
Li LiuJames E Strickler

Abstract

Recombinant protein expression in insect cells varies greatly from protein to protein. A fusion tag that is not only a tool for detection and purification, but also enhances expression and/or solubility would greatly facilitate both structure/function studies and therapeutic protein production. We have shown that fusion of SUMO (small ubiquitin-related modifier) to several test proteins leads to enhanced expression levels in Escherichia coli. In eukaryotic expression systems, however, the SUMO tag could be cleaved by endogenous desumoylase. In order to adapt SUMO-fusion technology to these systems, we have developed an alternative SUMO-derived tag, designated SUMOstar, which is not processed by native SUMO proteases. In the present study, we tested the SUMOstar tag in a baculovirus/insect cell system with several proteins, i.e. mouse UBP43, human tryptase beta II, USP4, USP15, and GFP. Our results demonstrate that fusion to SUMOstar enhanced protein expression levels at least 4-fold compared to either the native or His(6)-tagged proteins. We isolated active SUMOstar tagged UBP43, USP4, USP15, and GFP. Tryptase was active following cleavage with a SUMOstar specific protease. The SUMOstar system will make significant impact in di...Continue Reading

References

Apr 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·T R ButtS T Crooke
Feb 23, 1995·Nature·R HeimR Y Tsien
Jul 1, 1997·Biotechnology Progress·A PilonW E Bentley
Jan 24, 1998·The Journal of Biological Chemistry·C A GilchristR T Baker
Apr 2, 1998·Biological Chemistry·Z WangN M Schechter
Mar 27, 1999·Protein Expression and Purification·H ChanJ M Clark
Mar 27, 1999·Nature·S J Li, M Hochstrasser
Nov 5, 1999·The Journal of Biological Chemistry·A E ProudfootT N Wells
Mar 14, 2000·Molecular and Cellular Biology·S J Li, M Hochstrasser
Jun 1, 2000·Biochemical Society Transactions·R D PosseeL A King
Jul 25, 2000·Molecular & General Genetics : MGG·I SchwienhorstR J Dohmen
Jan 15, 2002·The Journal of Biological Chemistry·Michael P MalakhovDong-Er Zhang
May 16, 2002·Journal of Cellular Physiology·Keun Il KimChin Ha Chung
Jan 4, 2003·The Journal of Biological Chemistry·Allison M HunterPeter Liston
Jan 18, 2003·Mammalian Genome : Official Journal of the International Mammalian Genome Society·Corinne AngelatsRohan T Baker
Jun 25, 2003·Protein Expression and Purification·Tao WangDavid C H Yang
Sep 11, 2003·Biotechnology and Bioengineering·Luis MarangaManuel J T Carrondo
Apr 21, 2004·Protein Science : a Publication of the Protein Society·Ann-Maree CatanzaritiRohan T Baker
Jun 11, 2004·Annual Review of Biochemistry·Erica S Johnson
Jun 24, 2004·Seminars in Cell & Developmental Biology·Kenneth J Ritchie, Dong-Er Zhang
Jul 21, 2004·Journal of Structural and Functional Genomics·Michael P MalakhovTauseef R Butt
Oct 19, 2004·Journal of Virological Methods·Stephen A KabaMonique M van Oers
Oct 21, 2004·The Journal of Biological Chemistry·Tatiana A SobolevaRohan T Baker
Jan 26, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Sergey O TcherniukMaxim Y Balakirev
Aug 9, 2005·Protein Expression and Purification·Tauseef R ButtMichael R Mattern
Nov 9, 2005·Methods in Enzymology·Keun Il Kim, Dong-Er Zhang
Nov 9, 2005·Methods in Enzymology·Rohan T BakerPhilip G Board
Nov 22, 2005·The Journal of Biological Chemistry·Rob N de JongGert E Folkers
Dec 3, 2005·Protein Science : a Publication of the Protein Society·Jeffrey G MarblestoneTauseef R Butt
Dec 13, 2005·Molecular Pharmacology·Tetyana MilojevicChristian Nanoff
Aug 9, 2006·Protein Expression and Purification·Robert B KirkpatrickKyung Johanson
Feb 23, 2007·Current Pharmaceutical Design·C P Sommerhoff, N Schaschke
Feb 24, 2007·The EMBO Journal·Katrin SchweitzerMichael Naumann
Apr 22, 2008·Protein Science : a Publication of the Protein Society·Benjamin NicholsonTauseef R Butt

❮ Previous
Next ❯

Citations

Jun 24, 2009·Applied Microbiology and Biotechnology·Xianglei GaoSheng Xiong
Sep 10, 2013·Journal of Invertebrate Pathology·Saluana R CraveiroMaria Elita B Castro
Oct 20, 2010·Molecular & Cellular Proteomics : MCP·Christian M LochJames E Strickler
Jan 12, 2013·ACS Chemical Biology·Jennifer L BrighamDustin J Maly
Jul 22, 2014·Expert Opinion on Drug Discovery·Toshihiko SugikiChojiro Kojima
Sep 17, 2013·Protein Science : a Publication of the Protein Society·Mark R BellJames E Strickler
Dec 1, 2009·Biotechnology Journal·Artemisia M Andreou, Nektarios Tavernarakis
Nov 22, 2011·Protein Expression and Purification·Kathryn Rozen-GagnonSubhash G Vasudevan
Feb 16, 2010·Journal of Structural Biology·Joanne E NettleshipRaymond J Owens
Nov 5, 2016·Archives of Biochemistry and Biophysics·Dinesh K YadavNarendra Tuteja
Dec 22, 2017·The Journal of Cell Biology·Tino PleinerDirk Görlich
May 12, 2018·The Journal of Cell Biology·Metin AksuDirk Görlich
Feb 28, 2018·Nature Communications·Yibin XuMichael C Lawrence
Sep 4, 2020·Nature·Prashanth Srinivasan, Christina D Smolke
Apr 24, 2020·Nature·Susanne Hellmuth, Olaf Stemmann
Aug 23, 2020·Insect Biochemistry and Molecular Biology·Xien ChenSubba Reddy Palli
Jul 11, 2020·Cell Chemical Biology·Zachary E PotterDustin J Maly
Sep 29, 2011·Methods : a Companion to Methods in Enzymology·Richard B HitchmanLinda A King

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

© 2022 Meta ULC. All rights reserved