Translation levels control multi-spanning membrane protein expression.

PloS One
Hok Seon KimDaniel G Yansura

Abstract

Attempts to express eukaryotic multi-spanning membrane proteins at high-levels have been generally unsuccessful. In order to investigate the cause of this limitation and gain insight into the rate limiting processes involved, we have analyzed the effect of translation levels on the expression of several human membrane proteins in Escherichia coli (E. coli). These results demonstrate that excessive translation initiation rates of membrane proteins cause a block in protein synthesis and ultimately prevent the high-level accumulation of these proteins. Moderate translation rates allow coupling of peptide synthesis and membrane targeting, resulting in a significant increase in protein expression and accumulation over time. The current study evaluates four membrane proteins, CD20 (4-transmembrane (TM) helixes), the G-protein coupled receptors (GPCRs, 7-TMs) RA1c and EG-VEGFR1, and Patched 1 (12-TMs), and demonstrates the critical role of translation initiation rates in the targeting, insertion and folding of integral membrane proteins in the E. coli membrane.

References

Mar 1, 1978·Journal of Bacteriology·G F Miozzari, C Yanofsky
Mar 1, 1990·Trends in Genetics : TIG·J E McCarthy, C Gualerzi
Apr 10, 1987·Nucleic Acids Research·S Scholtissek, F Grosse
May 5, 1986·Journal of Molecular Biology·F W Studier, B A Moffatt
Jan 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·H A de BoerM Vasser
Sep 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·L W DeBoerE Braunwald
Jan 15, 1993·European Journal of Biochemistry·P R JensenO Michelsen
Nov 18, 1998·Annals of Oncology : Official Journal of the European Society for Medical Oncology·N L BerinsteinD Shen
Feb 4, 2005·Protein Science : a Publication of the Protein Society·Said EshaghiPär Nordlund
Apr 20, 2005·Current Opinion in Structural Biology·Pascal F EgeaPeter Walter
Sep 13, 2005·Annual Review of Microbiology·Joen LuirinkJan-Willem de Gier
Nov 16, 2005·Biochemistry·James A ErnstRichard L Vandlen
Jul 6, 2006·Trends in Biotechnology·Samuel WagnerJan-Willem de Gier
Apr 21, 2007·Molecular & Cellular Proteomics : MCP·Samuel WagnerJan-Willem de Gier
Aug 22, 2007·Proceedings of the National Academy of Sciences of the United States of America·Simon NewsteadDavid Drew
Sep 18, 2008·Proceedings of the National Academy of Sciences of the United States of America·Samuel WagnerJan-Willem de Gier
Sep 25, 2008·Proceedings of the National Academy of Sciences of the United States of America·Casim A SarkarAndreas Plückthun
May 12, 2009·Journal of the American Chemical Society·Linda ColumbusScott A Lesley

❮ Previous
Next ❯

Citations

Nov 10, 2013·Biochimica Et Biophysica Acta·Susan SchlegelJan-Willem de Gier
Jun 26, 2021·Microbial Biotechnology·Lisa Tietze, Rahmi Lale
Jul 3, 2019·ACS Synthetic Biology·Nico J ClaassensJohn van der Oost

❮ Previous
Next ❯

Methods Mentioned

BETA
density gradient centrifugation
pull-down
flow cytometry
FACS
size exclusion chromatography
ELISA
gel filtration
affinity purification

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.