ArticleThe Journal of biological chemistry2012
Green fluorescent protein changes the conductance of connexin 43 (Cx43) hemichannels reconstituted in planar lipid bilayers.
Article in The Journal of biological chemistry, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 28 citations in OpenAlex.
- Osteocyte connexin hemichannels and prostaglandin EProceedings of the National Academy of Sciences of the United States of America · 2025Article
- Overexpression of enhanced yellow fluorescent protein fused with Channelrhodopsin-2 causes contractile dysfunction in skeletal muscle.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2024Article
- SARS-CoV-2 spike protein S1 activates Cx43 hemichannels and disturbs intracellular CaBiological research · 2023Article
- Function Investigations and Applications of Membrane Proteins on Artificial Lipid Membranes.International journal of molecular sciences · 2023Review
- Cell-Free Protein Synthesis: A Promising Option for Future Drug Development.BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy · 2020Review
- Functional Reconstitution of Membrane Proteins Derived From Eukaryotic Cell-Free Systems.Frontiers in pharmacology · 2019Article
- Fluorescent dyes as a reliable tool in P2X7 receptor-associated pore studies.Journal of bioenergetics and biomembranes · 2015Article
- Growth arrest specific 1 (GAS1) is abundantly expressed in the adult mouse central nervous system.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2013Article
- Pannexin 1 ohnologs in the teleost lineage.The Journal of membrane biology · 2012Article
- Electrophysiology of single and aggregate Cx43 hemichannels.PloS one · 2012Article
- A microfluidic platform for measuring electrical activity across cells.Biomicrofluidics · 2012Article
Corrections and comments
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In mammalian tissues, connexin 43 (Cx43) is the most prominent member of the connexin family. In a single lipid bilayer, six connexin subunits assemble into a hemichannel (connexon). Direct communication of apposing cells is realized by two adjacent hemichannels, which can form gap junction channels. Here, we established an expression system in Pichia pastoris to recombinantly produce and purify Cx43 as well as Cx43 fused to green fluorescent protein (GFP). Proteins were isolated from crude cell membrane fractions via affinity chromatography. Cx43 and Cx43-GFP hemichannels were reconstituted in giant unilamellar vesicles as proven by fluorescence microscopy, and their electrophysiological behavior was analyzed on the single channel level by planar patch clamping. Cx43 and Cx43-GFP both showed an ohmic behavior and a voltage-dependent open probability. Cx43 hemichannels exhibited one major mean conductance of 224 ± 26 picosiemens (pS). In addition, a subconductance state at 124 ± 5 pS was identified. In contrast, the analysis of Cx43-GFP single channels revealed 10 distinct conductance states in the range of 15 to 250 pS, with a larger open probability at 0 mV as compared with Cx43, which suggests that intermolecular interactions between the GFP molecules alter the electrophysiology of the protein.
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