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    First product on the market for automated preparation of solvent-free giant unilamellar vesicles (GUVs)
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2009 - The M34A mutant of Connexin26 reveals active conductance states in pore-suspending membranes

icon pap  Port-a-Patch and   icon vpp   Vesicle Prep Pro publication in Journal of Structural Biology (2009)

Authors:
Gaßmann O., Kreir M., Ambrosi C., Pranskevich J., Oshima A, Röling C., Sosinsky G., Fertig N., Steinem C.

Journal: 
Journal of Structural Biology (2009) 168 (1):168-176


Abstract: 

Connexin26 (Cx26) is a member of the connexin family, the building blocks for gap junction intercellular channels. These dodecameric assemblies are involved in gap junction-mediated cell–cell communication allowing the passage of ions and small molecules between two neighboring cells. Mutations in Cx26 lead to the disruption of gap junction-mediated intercellular communication with consequences such as hearing loss and skin disorders. We show here that a mutant of Cx26, M34A, forms an active hemichannel in lipid bilayer experiments. A comparison with the Cx26 wild-type is presented. Two different techniques using micro/nano-structured substrates for the formation of pore-suspending lipid membranes are used. We reconstituted the Cx26 wild-type and Cx26M34A into artificial lipid bilayers and observed single channel activity for each technique, with conductance levels of around 35, 70 and 165 pS for the wild-type. The conductance levels of Cx26M34A were found at around 45 and 70 pS.


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