A method is described that uses trypsin digestion combined with collagenase-hyaluronidase which produces a population of gap junction vesicles. The hexagonal lattice of subunits ("connexons") comprising the gapjunctions appears unaltered by various structural criteria and by buoyant density measurements. The gap junction vesciles are closed by either a single or a double profile of nonjunctional "membrane," which presents a smooth, particle-free fracture face. Horseradish peroxidase and cytochrome c studies have revealed that about 20% of the gap junction vesicles are impermeable to proteins 12,000 daltons or larger. The increased purity of the trypsinized junction preparation suggests that one of the disulfide reduction products of the gap-junction principal protein may be a nonjunctional contaminating peptide. The gap junction appears to be composed of a single 18,000-dalton protein, connexin, which may be reduced to a single 9,000-dalton peak. The number of peptides in this reduced peak are still unknown.
Article| February 01 1976
In vitro formation of gap junction vesicles.
D A Goodenough
Online Issn: 1540-8140
Print Issn: 0021-9525
J Cell Biol (1976) 68 (2): 220–231.
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D A Goodenough; In vitro formation of gap junction vesicles.. J Cell Biol 1 February 1976; 68 (2): 220–231. doi: https://doi.org/10.1083/jcb.68.2.220
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