Coin J T, Olson J S
J Biol Chem. 1979 Feb 25;254(4):1178-90.
Oxygen uptake into intact and reconstituted human red blood cells was measured using dual wavelength, stopped flow techniques. The rate of oxygen uptake by human erythrocytes is roughly 40 times slower (t 1/2 congruent to 80 ms at 0.125 mM O2, 25 degrees C) than the corresponding rate of oxygen combination with free hemoglobin. Oxygen transport through the red cell cytoplasm accounts for part of this difference and predicts a half-time of uptake of about 15 ms, which is still 5 times smaller than that observed experimentally. Further limitation of uptake appears to be due to the presence of unstirred layers of solvent adjacent to the red cell surface. Very rapidly after mixing, these layers form and become depleted of O2 due to uptake by the cells. This requires that the bulk of the oxygen molecules must diffuse over rather large distances, 1.0 to 5.0 micrometer, before they can penetrate the erythrocytes. A mathematical model was developed to take into account diffusion through an unstirred solvent layer which increases in thickness with time. This scheme can account quantitatively both for the dependence of the apparent rate of uptake on O2 concentration and for the shape of the observed time courses. In addition, the diffusion parameters which were developed for the O2 reaction can also be used to describe quantitatively the rates and time courses of CO and ethyl isocyanide uptake and the rates and time courses of O2 release from cells in the presence of sodium dithionite. Finally, the parameters used to describe the stopped flow results can also be used to simulate quantitatively O2 uptake time courses obtained from previous studies with thin films of red cells (Sinha, A. K. (1969) Ph.D. dissertation, University of California, San Francisco; Thews, G. (1959) Arch. Gesamte Physiol. Mens. Tiere (Pflufgers) 268, 308-317).
采用双波长停流技术测量完整的和重构的人红细胞对氧气的摄取。人红细胞摄取氧气的速率大约比游离血红蛋白与氧气结合的相应速率慢40倍(在0.125 mM氧气、25℃时,t1/2约为80毫秒)。氧气通过红细胞细胞质的运输是造成这种差异的部分原因,预计摄取的半衰期约为15毫秒,这仍比实验观察到的结果小5倍。摄取的进一步限制似乎是由于红细胞表面附近存在未搅动的溶剂层。混合后很快,这些层就形成了,并且由于细胞摄取氧气而使氧气耗尽。这就要求大部分氧分子在穿透红细胞之前必须扩散相当大的距离,即1.0至5.0微米。建立了一个数学模型来考虑通过随时间增加厚度的未搅动溶剂层的扩散。该模型既能定量解释摄取的表观速率对氧气浓度的依赖性,又能解释观察到的时间进程的形状。此外,为氧气反应建立的扩散参数也可用于定量描述一氧化碳和乙基异氰化物摄取的速率和时间进程,以及在连二亚硫酸钠存在下细胞释放氧气的速率和时间进程。最后,用于描述停流结果的参数也可用于定量模拟先前用红细胞薄膜研究得到的氧气摄取时间进程(辛哈,A.K.(1969年),博士论文,加利福尼亚大学旧金山分校;特威斯,G.(1959年),《人体及动物生理学文献集(普弗吕格尔斯)》268,308 - 317)。