Connexins and the vacuolar proteolipid-like 16-kDa protein are not directly associated with each other but may be components of similar or the same gap junctional complexes.
Gap junction preparations made from mouse liver plasma membranes by alkali extraction contain variable proportions of connexins (Cx32 and Cx26) and the 16-kDa protein which is closely related or may be identical to the 16-kDa proteolipid (subunit c) of the vacuolar H(+)-ATPase and the mediatophore complex. The absence of a stoichiometric relationship suggests that connexins and the 16-kDa protein are not subunits of the same channel complex, but analysis of alkali preparations by isopycnic centrifugation shows both types of protein are in membrane structures of the same buoyant density. Electron microscopic analysis of alkali preparations shows a homogeneous population of gap junctions of uniform morphology and width, suggesting the proteins are in the same or similar structures. The structures containing connexins and the 16-kDa protein can be separated by treatment of the plasma membranes with Triton X-100. After such treatment, the connexins remain associated with dense cellular or extracellular material and the gap junctional structures, after further extraction with N-lauroyl sarcosine and urea, contain only the 16-kDa protein. These detergent-extracted gap junctions are thinner (14.1 nm) than those in alkali preparations (18.4 nm).
Finbow
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ME
CRC Laboratories, Beatson Institute for Cancer Research, Garscube Estate, Glasgow, Scotland.
Meagher
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D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000648:ultrastructure; D017630:Connexins; D003902:Detergents; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D007365:Intercellular Junctions / Q000737:chemistry* / Q000648:ultrastructure*; D008099:Liver / Q000737:chemistry* / Q000648:ultrastructure*; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000032:analysis* / Q000302:isolation & purification; D051379:Mice; D008854:Microscopy, Electron; D008970:Molecular Weight; D017830:Octoxynol; D010449:Peptide Mapping; D011092:Polyethylene Glycols; D011510:Proteolipids / Q000032:analysis*; D006180:Proton-Translocating ATPases / Q000032:analysis; D014617:Vacuoles / Q000737:chemistry* / Q000648:ultrastructure*
D017630:Connexins; D003902:Detergents; D046911:Macromolecular Substances; D008565:Membrane Proteins; D011510:Proteolipids; D011092:Polyethylene Glycols; D017830:Octoxynol; D006180:Proton-Translocating ATPases