Miller D S, Horowitz S B
J Biol Chem. 1986 Oct 25;261(30):13911-5.
The intracellular distribution and diffusivity of adenosine triphosphate (ATP) was studied by cryomicrodissection of individual Rana pipiens oocytes. We measured ATP concentrations in the nucleus, in animal and vegetal hemisphere cytoplasm, and in an intracellular reference phase (iRP, a microinjected gelatin "organelle") which samples diffusive ATP. Regional concentrations were not equal: nucleus much greater than animal ooplasm greater than vegetal ooplasm. ATP binding and water availability (as solvent) were determined by plotting nuclear and cytoplasmic ATP concentrations as a function of reference phase ATP concentrations (isothermal analysis). The nucleus/iRP isotherm for ATP was an equimolar line, showing that nucleoplasm resembles iRP gelatin (and consequently a simple aqueous solution) in its solvent properties. Cytoplasm/iRP isotherms were more complex, having slopes much less than unity and ordinal intercepts above the graph's origin. They demonstrate the presence in cytoplasm of mechanisms that are capable of excluding and binding ATP. These mechanisms are responsible for the inhomogeneity in ATPs intracellular distribution. In addition, exclusion and binding have different and opposing effects on ATP concentrations in the cell's "soluble space," and hence on ATP availability to enter into cellular reactions. It follows that these phenomena must be considered in attempts to model ATPs role in metabolism.
通过对单个牛蛙卵母细胞进行冷冻显微切割,研究了三磷酸腺苷(ATP)的细胞内分布和扩散率。我们测量了细胞核、动物半球和植物半球细胞质以及细胞内参考相(iRP,一种显微注射的明胶“细胞器”,用于采样扩散的ATP)中的ATP浓度。区域浓度并不相等:细胞核中的浓度远高于动物卵质,动物卵质又高于植物卵质。通过绘制细胞核和细胞质ATP浓度作为参考相ATP浓度的函数图(等温分析),确定了ATP结合和水的可利用性(作为溶剂)。ATP的细胞核/iRP等温线是一条等摩尔线,表明核质在其溶剂性质上类似于iRP明胶(因此是一种简单的水溶液)。细胞质/iRP等温线更为复杂,斜率远小于1,且有序截距在图的原点上方。它们表明细胞质中存在能够排斥和结合ATP的机制。这些机制导致了ATP细胞内分布的不均匀性。此外,排斥和结合对细胞“可溶空间”中的ATP浓度有不同且相反的影响,从而对ATP参与细胞反应的可用性产生影响。因此,在试图模拟ATP在代谢中的作用时,必须考虑这些现象。