| Single channel behavior of recombinant beta 2 gap junction connexons reconstituted into planar lipid bilayers Biophysical Journal, Volume 68, Issue 5, 1 May 1995, Pages 1767-1775 L.K. Buehler, K.A. Stauffer, N.B. Gilula and N.M. Kumar Abstract The beta 2 gap junction protein (Cx26) was expressed in an insect cell line by infection with a baculovirus vector containing the rat beta 2 cDNA. Isolated beta 2 gap junction connexons were reconstituted into planar lipid bilayers. Single channel activity was observed with a unitary conductance of 35–45 pS in 200 mM KCl. Channels with conductance values of 60 pS and 90–110 pS also coexisted with the lower conducting channel suggesting that there are channels with different conductance properties within a population of connexons. Channel activity was observed at voltages of up to 150 mV. Furthermore, the characterization of these channel properties from the beta 2 connexons that were generated by this heterologous expression system has provided the basis for identifying an endogenous beta 2 connexon channel in material reconstituted from native rat liver gap junctions. Abstract | PDF (1355 kb) |
| The Gap Junction Communication Channel Cell, Volume 84, Issue 3, 9 February 1996, Pages 381-388 Nalin M Kumar and Norton B Gilula Full Text | PDF (921 kb) |
| Correlation analysis of gap junction lattice images Biophysical Journal, Volume 58, Issue 5, 1 November 1990, Pages 1213-1226 G.E. Sosinsky, T.S. Baker, D.L. Caspar and D.A. Goodenough Abstract Fourier averages of connexon images computed from low-irradiation electron micrographs of isolated negatively stained gap junction domains exhibited differences in stain distribution and connexon orientation. To analyze these polymorphic structures, correlation averaging methods were applied to images from negatively stained and frozen-hydrated specimens. For the negatively stained specimens, separate averages over two subsets of connexons with differing degrees of stain accumulation in the axial channel were obtained. Two populations of connexons with opposite skew orientations were distinguishable within a single junctional domain of a frozen-hydrated specimen. Correlation maps calculated using the left- and right-skewed references showed that the selected connexons tend to locally cluster. Using correlation methods to analyze packing disorder in a typical connexon lattice, we estimated the root-mean-square variation in the nearest neighbor pair separation to be approximately 11% of the lattice constant. Displacements of the connexons relative to each other increased with increasing pair separation in the lattice, rather like a liquid, although long-range orientation order was conserved as in a crystal. These results support the hypothesis that the hexagonal ordering of the connexons results from short-range repulsive forces. Abstract | PDF (4602 kb) |
Copyright © 1984 The Biophysical Society. All rights reserved.
Biophysical Journal, Volume 45, Issue 1, 208-218, 1 January 1984
doi:10.1016/S0006-3495(84)84149-1
Research Article
L. Makowski, D.L. Caspar, W.C. Phillips, T.S. Baker and D.A. Goodenough
Correlation of structural changes in isolated gap junctions with the mechanism of channel gating is complicated by the effects of isolation procedures and the lack of a direct functional assay. The effect of variations in the isolation procedure are examined by comparison of the structures of gap junctions isolated by different protocols. X-ray diffraction data from over two hundren specimens are compared to provide a basis for identification of invariant aspects of the connexon structure and variable properties related either to functional switching or experimental modifications. We discuss the relationship between subunit tilt, lattice symmetry and packing, and membrane curvature and demonstrate that membrane curvature may be a natural consequence of the structure of the connexons and the patterns of interactions between them.