Cell-free synthesis and assembly of connexins into functional gap junction membrane channels

The EMBO Journal
M M FalkN B Gilula

Abstract

Several different gap junction channel subunit isotypes, known as connexins, were synthesized in a cell-free translation system supplemented with microsomal membranes to study the mechanisms involved in gap junction channel assembly. Previous results indicated that the connexins were synthesized as membrane proteins with their relevant transmembrane topology. An integrated biochemical and biophysical analysis indicated that the connexins assembled specifically with other connexin subunits. No interactions were detected between connexin subunits and other co-translated transmembrane proteins. The connexins that were integrated into microsomal vesicles assembled into homo- and hetero-oligomeric structures with hydrodynamic properties of a 9S particle, consistent with the properties reported for hexameric gap junction connexons derived from gap junctions in vivo. Further, cell-free assembled homo-oligomeric connexons composed of beta1 or beta2 connexin were reconstituted into synthetic lipid bilayers. Single channel conductances were recorded from these bilayers that were similar to those measured for these connexons produced in vivo. Thus, this is the first direct evidence that the synthesis and assembly of a gap junction connexo...Continue Reading

References

Nov 12, 1992·Nature·R L Rosenberg, J E East
Oct 1, 1991·The Journal of Cell Biology·K A StaufferN Unwin
Jan 1, 1985·The Journal of Membrane Biology·R WernerG Dahl
Jan 1, 1989·Annual Review of Cell Biology·S M Hurtley, A Helenius
Oct 22, 1987·Nature·B NicholsonJ P Revel
Sep 25, 1984·Nucleic Acids Research·P A Krieg, D A Melton
Feb 7, 1980·Nature·P N Unwin, G Zampighi
Aug 4, 1995·The Journal of Biological Chemistry·M CascioN B Gilula
Oct 1, 1994·The Journal of Cell Biology·M M FalkN B Gilula
Aug 1, 1995·Microscopy Research and Technique·M Yeager
Feb 6, 1996·Proceedings of the National Academy of Sciences of the United States of America·J X Jiang, D A Goodenough
Feb 9, 1996·Cell·N M Kumar, N B Gilula
Apr 1, 1996·Current Opinion in Structural Biology·M Yeager, B J Nicholson
Jan 1, 1996·Annual Review of Biochemistry·D A GoodenoughD L Paul
Jan 1, 1997·Nature Structural Biology·V M UngerM Yeager

❮ Previous
Next ❯

Citations

Aug 26, 1999·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·R J StockertE L Hertzberg
Jun 30, 2000·Cell Biology International·G J Snipes, W Orfali
Jul 28, 2007·The Journal of Membrane Biology·Yasufumi OmoriKatsuhiko Enomoto
Jun 29, 2012·The Journal of Membrane Biology·Scott R JohnstoneBrant E Isakson
Sep 28, 2007·Applied Microbiology and Biotechnology·Haiqin ChenPeilin Cen
Apr 7, 2000·Brain Research. Brain Research Reviews·C Ressot, R Bruzzone
Nov 20, 1998·Current Opinion in Biotechnology·L JermutusA Plückthun
Sep 8, 1998·Current Opinion in Structural Biology·M YeagerM M Falk
May 6, 2010·European Journal of Human Genetics : EJHG·Ching-Chyuan SuJiann-Jou Yang
Feb 11, 2012·Proceedings of the National Academy of Sciences of the United States of America·Carmen E FloresAlberto E Pereda
May 4, 2011·The Journal of Biological Chemistry·Alfredo G FortDavid C Spray
Oct 1, 2005·Molecular Biology of the Cell·Andrew W HunterRobert G Gourdie
Jan 27, 2006·Molecular Biology of the Cell·Corinna LehmannMichael Hoch
Mar 20, 2009·Molecular Biology of the Cell·Judy K VanslykeLinda S Musil
Jan 13, 2010·Cold Spring Harbor Perspectives in Biology·Daniel A Goodenough, David L Paul
Nov 24, 2004·Clinical and Experimental Pharmacology & Physiology·Nicole M Rummery, Caryl E Hill
Apr 25, 2012·Circulation·Sakthivel VaiyapuriJonathan M Gibbins
May 23, 2001·Archives of Biochemistry and Biophysics·P D Lampe, A F Lau
Aug 1, 2002·Proceedings of the National Academy of Sciences of the United States of America·Undine LaufMatthias M Falk
Jan 27, 2010·Otology & Neurotology : Official Publication of the American Otological Society, American Neurotology Society [and] European Academy of Otology and Neurotology·Susan ArndtRalf Birkenhäger
Jun 22, 2012·Molecular Membrane Biology·Christian RoosFrank Bernhard
Jun 18, 2002·Cell Communication & Adhesion·G Richard
Jun 18, 2002·Cell Communication & Adhesion·P LopezS C Chen
Feb 25, 2009·Journal of Structural Biology·Oliver GassmannClaudia Steinem
May 14, 2004·The Journal of Investigative Dermatology·Johanna M BrandnerIngrid Moll
Nov 17, 2007·The Journal of Investigative Dermatology·Joey E Lai-CheongJohn A McGrath
Sep 4, 2008·Proteomics·Daniel SchwarzFrank Bernhard
Jan 4, 2006·Biochimica Et Biophysica Acta·Carsten ZeilingerHans-Albert Kolb

❮ 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.