Horowitz S B, Fenichel I R, Hoffman B, Kollmann G, Shapiro B
Biophys J. 1970 Oct;10(10):994-1010. doi: 10.1016/S0006-3495(70)86348-2.
Radioautography and extractive techniques were used to analyze the transport of cysteamine phosphate and its derivatives in salamander oocytes. The quantitative relations among the processes involved - membrane permeation, enzymatic dephosphorylation, binding through mixed disulfide formation, and cytoplasmic diffusion - were elucidated. Within the detection limits, all of the intracellular material is present as dephosphorylated derivatives. Cytoplasmic diffusion is effectively slowed by binding (the "chromatographic" effect) and makes an appreciable contribution to cellular flux rates. As a consequence, one can observe by radioautography a cortical diffusion ring which spreads inward as a function of influx time, while also increasing in peak density because of the finite membrane permeability. Good agreement was found between the transport parameters determined by radioautography and those from influx data for the whole oocyte. The ratio of nuclear to cytoplasmic concentrations of the cysteamine phosphate derivatives at equilibrium is about 0.4. The nuclear membrane is, however, a negligible barrier to transport, and the asymmetry appears to arise primarily from the quantity and sulfhydryl content of the binding proteins in the two compartments.
利用放射自显影术和提取技术分析了磷酸半胱胺及其衍生物在蝾螈卵母细胞中的转运。阐明了所涉及过程之间的定量关系——膜渗透、酶促去磷酸化、通过混合二硫键形成的结合以及细胞质扩散。在检测限内,所有细胞内物质均以去磷酸化衍生物的形式存在。结合作用(“色谱”效应)有效地减缓了细胞质扩散,并对细胞通量速率做出了显著贡献。因此,通过放射自显影可以观察到一个皮质扩散环,它随着流入时间向内扩散,同时由于有限的膜通透性,其峰值密度也会增加。放射自显影测定的转运参数与整个卵母细胞流入数据确定的参数之间发现了良好的一致性。平衡时,磷酸半胱胺衍生物的核浓度与细胞质浓度之比约为0.4。然而,核膜对转运的屏障作用可忽略不计,这种不对称性似乎主要源于两个区室中结合蛋白的数量和巯基含量。