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缺乏三磷酸腺苷的卡尔斯伯酵母制剂中甘氨酸的浓度:质子梯度和解偶联剂的影响

The concentration of glycine by preparations of the yeast Saccharomyces Carlsbergensis depleted of adenosine triphosphate: Effects of proton gradients and uncoupling agents.

作者信息

Seaston A, Carr G, Eddy A A

出版信息

Biochem J. 1976 Mar 15;154(3):669-76. doi: 10.1042/bj1540669.

Abstract
  1. At pH 4.5 and 30degreesC, yeast preparations depleted of ATP in the presence of antimycin and deoxyglucose spontaneously lost K+, gaining roughly an equivalent amount of H+. 2. Five proton conductors including azide and 2,4-dinitrophenol accelerated this process, as did [14C]glycine, which was absorbed with two extra equivalents of H+. 3. The rate of glycine uptake at pH 4.5 diminished fourfold when cellular K+ fell by 20%. 4. The distribution of [14C]propionate indicated that the intracellular pH fell from 6.2 to 5.7 when the cellular content of K+ fell by 30%. 5. Glycine uptake from a 5 muM solution was about 400 times faster at pH 4.5 than it was at pH 7.4 with 100mM-KC1 present ostensibly to lower the membrane potential. 6. Yeast preparations containing 2mM-[14C]glycine absorbed a further amount from a 0.1 muM solution at pH 4.5. After about 10 min a net movement of [14C]glycine out of the yeast occurred. The ratio of the cellular [14Ia1glycine concentration to the concentration outside the yeast reached 4 X 10(4) in these assays, whereas at pH 7.4 in the presence of 100mM-KC1 it did not exceed 15 in 3h. Dimitrophenol lowered the accumulation ratio at pH 4.5, apparently by causing proton conduction. 7. The observations are consistent with the notion that glycine uptake is driven by a proton symport mechanism. 8. Possible factors governing the strikingly low rate of glycine efflux as opposed to its optimum rate of influx are discussed.
摘要
  1. 在pH值为4.5、温度为30摄氏度的条件下,在抗霉素和脱氧葡萄糖存在时耗尽ATP的酵母制剂会自发失去钾离子,同时获得大致等量的氢离子。2. 包括叠氮化物和2,4-二硝基苯酚在内的五种质子导体加速了这一过程,[14C]甘氨酸也有同样作用,它吸收时会额外伴随两当量的氢离子。3. 当细胞内钾离子含量下降20%时,pH值为4.5时甘氨酸的摄取速率降低了四倍。4. [14C]丙酸盐的分布表明,当细胞内钾离子含量下降30%时,细胞内pH值从6.2降至5.7。5. 在pH值为4.5时,从5μM溶液中摄取甘氨酸的速度比在pH值为7.4且存在100mM - KCl(表面上是为了降低膜电位)时快约400倍。6. 含有2mM - [14C]甘氨酸的酵母制剂在pH值为4.5时从0.1μM溶液中进一步吸收甘氨酸。大约10分钟后[14C]甘氨酸开始从酵母中净流出。在这些实验中,酵母细胞内[14C]甘氨酸浓度与酵母外浓度的比值达到4×10⁴,而在pH值为7.4且存在100mM - KCl时,3小时内该比值不超过15。二硝基苯酚降低了pH值为4.5时的积累比值,显然是通过引起质子传导实现的。7. 这些观察结果与甘氨酸摄取由质子同向转运机制驱动这一观点一致。8. 讨论了与甘氨酸最佳流入速率相比,导致其流出速率极低的可能因素。

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