Stephens R E
J Cell Biol. 1975 Feb;64(2):408-20. doi: 10.1083/jcb.64.2.408.
The basal apparatus, consisting of an array of interconnected basal bodies bearing bifurcating striated rootlets encompassing a nucleus, has been isolated from hypertonically deciliated columnar gill epithelial cells of the bay scallop Aequipecten irradians through gentle lysis with Triton X-100. The rootlets, 8-10 mum in length, were not easily preserved with conventional electron microscope fixatives, suggesting that the extent of their contribution to cellular architecture has been somewhat underestimated, even though Englemann described many of the structural details of the basal apparatus in 1880. The striated rootlets were soluble at high but not at low pH, in 2 M solutions of sodium azide and potassium thiocyanate but not sodium or potassium chloride, in 1% deoxycholate but not digitonin, and in the denaturing solvents 6 M guanidine-HC1, 8 M urea, and 1% sodium dodecylsulfate at 100 degrees C. The protein found consistently when rootlets were solubilized migrated on SDS-polyacrylamide gels as a closely spaced doublet with apparent molecular weights of 230,000 and 250,000 daltons. This unique protein, distinct from tropocollagen or various muscle components, has been named ankyrin because of the rootlet's anchor-like function in the cell.
通过用Triton X-100温和裂解,从海湾扇贝栉孔扇贝经高渗去纤毛的柱状鳃上皮细胞中分离出了基部装置,该装置由一系列相互连接的基体组成,这些基体带有包围细胞核的分叉横纹小根。这些小根长度为8-10微米,用传统的电子显微镜固定剂不易保存,这表明即使恩格尔曼在1880年描述了基部装置的许多结构细节,它们对细胞结构的贡献程度在一定程度上仍被低估。横纹小根在高pH但低pH下可溶,在2M叠氮化钠和硫氰酸钾溶液中可溶但在氯化钠或氯化钾溶液中不溶,在1%脱氧胆酸盐中可溶但在洋地黄皂苷中不溶,在变性溶剂6M盐酸胍、8M尿素和1%十二烷基硫酸钠中于100℃时可溶。当小根溶解时始终能发现的蛋白质在SDS-聚丙烯酰胺凝胶上迁移为紧密间隔的双峰,表观分子量分别为230,000和250,000道尔顿。这种独特的蛋白质不同于原胶原蛋白或各种肌肉成分,由于小根在细胞中的锚定样功能而被命名为锚蛋白。