GAP JUNCTION

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Gap junctions (GJs) represent a ubiquitous component of all multicellular animals, and GJ s were one of the first specimens examined by electron crystallographic methods. However, the majority (99.9%) of these crystals have good short range order combined with long range disorder that results in coherent diffraction data generally only to about ~15-25 ¿. As a result, there is only one 3D structure obtained at > 10 ¿ (Unger et al., 1999). This is an important point because one of the aims is to reinvent image processing procedures that would allow us to obtain structural data based on a hybrid crystallographic/single particle reconstruction approach for greater throughput. Furthermore, we have found that there are classes of interesting connexin functional mutants where the hexamer or dodecamer is stable when held within the membrane, but dissociates upon detergent solubilization. For these mutants, crystallization for a high resolution structure will never be possible. We began development of hybrid methods of image processing combining crystallographic information with single particle reconstruction methods so it is much less dependent on lattice order.