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卡尔酵母(Saccharomyces carlsbergensis)吸收氨基酸时伴随的化学计量质子和钾离子运动。

Stoicheiometrical proton and potassium ion movements accompanying the absorption of amino acids by the yeast Saccharomyces carlsbergensis.

作者信息

Eddy A A, Nowacki J A

出版信息

Biochem J. 1971 May;122(5):701-11. doi: 10.1042/bj1220701.

Abstract
  1. Proton uptake into the yeast Saccharomyces carlsbergensis, was studied at pH4.5-5.5 in the presence of both antimycin and 2-deoxyglucose to inhibit energy metabolism. Previous work had shown that the cells then absorbed about 20nmol of glycine or l-phenylalanine against a considerable amino acid concentration gradient. The addition of the amino acid immediately stimulated the rate of uptake of protons two- to three-fold. About 2 extra equivalents of H(+) accompanied a given amount of the amino acids into the yeast preparations exposed to the metabolic inhibitors for 2-4min and about 1.2 equivalents after 20min exposure. 2. Analogous observations were made during serial additions of glycine, l-phenylalanine, l-leucine and l-lysine to preparations lacking the metabolic inhibitors and deficient in substrates needed for energy metabolism. In fresh cellular preparations the influx of glycine was then closely coupled to a stimulated flow of 2.1 equiv. of H(+) into the yeast. A similar number of K(+) ions left the cells. About 30% of the extra protons was subsequently ejected from the yeast. Deoxyglucose and antimycin together inhibited the ejection of protons. When the yeast had been fed with glucose energy metabolism was stimulated and almost as many protons as were absorbed with the amino acid were apparently ejected again. 3. Yeast preparations containing Na(+), instead of K(+), as the principal cation absorbed about 1 extra equivalent of H(+) after the addition of phenylalanine, glycine or leucine. This response was not observed in the presence of both deoxyglucose and antimycin. 4. The observations show that H(+) and, in certain circumstances, K(+) are co-substrates in the transport of the amino acids into the yeast. An analogy is drawn with the roles of Na(+) and K(+) as co-substrates in certain mammalian systems. The results lead to various models relating the physical flow of the co-substrate ions on the amino acid carrier to the transduction of chemical energy in an associated ion pump forming part of the mechanism for transporting amino acids into the yeast.
摘要
  1. 在抗霉素和2-脱氧葡萄糖存在的情况下,于pH4.5 - 5.5研究了质子进入卡尔酵母(Saccharomyces carlsbergensis)的过程,以抑制能量代谢。先前的研究表明,此时细胞会逆着相当大的氨基酸浓度梯度吸收约20纳摩尔的甘氨酸或L-苯丙氨酸。氨基酸的添加立即使质子吸收速率提高了两到三倍。在暴露于代谢抑制剂2 - 4分钟的酵母制剂中,每给定数量的氨基酸会伴随约2额外当量的H⁺进入,暴露20分钟后约为1.2当量。2. 在向缺乏代谢抑制剂且能量代谢所需底物不足的制剂中连续添加甘氨酸、L-苯丙氨酸、L-亮氨酸和L-赖氨酸的过程中,也进行了类似的观察。在新鲜的细胞制剂中,甘氨酸的流入与2.1当量的H⁺刺激流入酵母紧密相关。类似数量的K⁺离子离开细胞。随后约30%的额外质子从酵母中排出。脱氧葡萄糖和抗霉素共同抑制质子的排出。当酵母用葡萄糖喂养时,能量代谢受到刺激,并且几乎与氨基酸吸收时吸收的质子数量相同的质子显然又被排出。3. 以Na⁺而非K⁺作为主要阳离子的酵母制剂在添加苯丙氨酸、甘氨酸或亮氨酸后吸收了约1额外当量的H⁺。在脱氧葡萄糖和抗霉素同时存在的情况下未观察到这种反应。4. 这些观察结果表明,H⁺以及在某些情况下K⁺是氨基酸转运进入酵母过程中的共底物。与Na⁺和K⁺在某些哺乳动物系统中作为共底物的作用进行了类比。结果产生了各种模型,将共底物离子在氨基酸载体上的物理流动与相关离子泵中化学能的转导联系起来,该离子泵构成了氨基酸转运进入酵母机制的一部分。

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