Kistler J, Bond J, Donaldson P, Engel A
School of Biological Sciences, Centre for Gene Technology, University of Auckland, New Zealand.
J Struct Biol. 1993 Jan-Feb;110(1):28-38. doi: 10.1006/jsbi.1993.1002.
The in vitro assembly of crystalline gap junctions from detergent-solubilized sheep lens fiber cell pore complexes in the presence of endogenous lipid is a two-step process. "Mini"-gap junctions containing about 10 pore complexes assembled within 12 hr of dialysis. The removal of detergent appeared to be the sole factor required to reach this level of assembly. Further growth to micrometer-sized crystalline gap junctions required cleaved connexin, MgCl2, elevated dialysis temperature, and dialysis for 3-5 days. Molecular interactions between adjacent pore structures laterally in the membrane are vital for the second phase of assembly but appear to be of minor importance for the formation of mini-gap junctions. Parallels exist between structural aspects of gap junction assembly in vitro and the 2-D crystallization of membrane proteins in general.
在存在内源性脂质的情况下，从经去污剂溶解的绵羊晶状体纤维细胞孔复合物进行晶体间隙连接的体外组装是一个两步过程。含有约10个孔复合物的“微型”间隙连接在透析12小时内组装完成。去污剂的去除似乎是达到这一组装水平所需的唯一因素。进一步生长为微米级晶体间隙连接需要切割的连接蛋白、MgCl2、提高透析温度以及透析3至5天。膜中相邻孔结构之间的分子相互作用对组装的第二阶段至关重要，但对微型间隙连接的形成似乎不太重要。体外间隙连接组装的结构方面与一般膜蛋白的二维结晶之间存在相似之处。
