Carruthers A, Melchior D L
Biochemistry. 1985 Jul 16;24(15):4244-50. doi: 10.1021/bi00336a065.
The kinetics of alpha- and beta-D-glucose mutarotation and the transport of these anomers by intact human red cells were determined at 0.6 and 36.6 degrees C. The mutarotation coefficients for alpha- and beta-D-glucose in cell-free tris(hydroxymethyl)aminomethane medium (pH 7.4) at 0.6 degrees C are (2.25 +/- 0.2) and (1.73 +/- 0.42) X 10(-3) min-1, respectively, and at 36.6 degrees C are (69 +/- 12) and (75 +/- 5) X 10(-3) min-1, respectively. These values are in good agreement with previous estimates. At 0.6 degrees C, the red cell contains no detectable mutarotase activity. Initial rates of sugar uptake were measured by using radiolabeled D-glucose and time courses of uptake by turbidimetry. The time courses of alpha- and beta-D-glucose and an equilibrium mixture of alpha- and beta-D-glucose infinite-cis entry are identical at 0.66 degrees C (n = 41) where negligible mutarotation is observed. The apparent Ki values for inhibition of radiolabeled D-glucose initial uptake by unlabeled alpha- or beta-D-glucose at 0.6 degrees C are identical (1.6 mM). The calculated Vmax parameters for uptake of the radiolabeled anomers at this temperature are also indistinguishable. The time courses of infinite-cis alpha- and beta-D-glucose uptake at 36.66 degrees C are identical (n = 40). While D-glucose mutarotation is more rapid at this temperature, the anomers of D-glucose are not transported differently by the red cell hexose transfer system.(ABSTRACT TRUNCATED AT 250 WORDS)
在0.6℃和36.6℃下测定了α - 和β - D - 葡萄糖变旋的动力学以及这些异头物通过完整人类红细胞的转运情况。在0.6℃时,无细胞的三(羟甲基)氨基甲烷培养基(pH 7.4)中α - 和β - D - 葡萄糖的变旋系数分别为(2.25±0.2)和(1.73±0.42)×10⁻³ min⁻¹;在36.6℃时分别为(69±12)和(75±5)×10⁻³ min⁻¹。这些值与先前的估计值吻合良好。在0.6℃时,红细胞中未检测到变旋酶活性。通过使用放射性标记的D - 葡萄糖测量糖摄取的初始速率,并通过比浊法测定摄取的时间进程。在0.66℃(n = 41)时,α - 和β - D - 葡萄糖以及α - 和β - D - 葡萄糖无限顺式进入的平衡混合物的摄取时间进程相同,此时观察到的变旋可忽略不计。在0.6℃时,未标记的α - 或β - D - 葡萄糖对放射性标记的D - 葡萄糖初始摄取的抑制作用的表观Ki值相同(1.6 mM)。在此温度下,计算得到的放射性标记异头物摄取的Vmax参数也无差异。在36.66℃时,无限顺式α - 和β - D - 葡萄糖摄取的时间进程相同(n = 40)。虽然在此温度下D - 葡萄糖变旋更快,但红细胞己糖转运系统对D - 葡萄糖的异头物转运并无差异。(摘要截短于250字)