Russell J B, Strobel H J, Driessen A J, Konings W N
U.S. Department of Agriculture, Cornell University, Ithaca, New York 14853.
J Bacteriol. 1988 Aug;170(8):3531-6. doi: 10.1128/jb.170.8.3531-3536.1988.
Streptococcus bovis JB1 cells were able to transport serine, threonine, or alanine, but only when they were incubated in sodium buffers. If glucose-energized cells were washed in potassium phosphate and suspended in potassium phosphate buffer, there was no detectable uptake. Cells deenergized with 2-deoxyglucose and incubated in sodium phosphate buffer were still able to transport serine, and this result indicated that the chemical sodium gradient was capable of driving transport. However, when the deenergized cells were treated with valinomycin and diluted into sodium phosphate to create both an artificial membrane potential and a chemical sodium gradient, rates of serine uptake were fivefold greater than in cells having only a sodium gradient. If deenergized cells were preloaded with sodium (no membrane potential or sodium gradient), there was little serine transport. Nigericin and monensin, ionophores capable of reversing sodium gradients across membranes, strongly inhibited sodium-dependent uptake of the three amino acids. Membrane vesicles loaded with potassium and diluted into either lithium or choline chloride were unable to transport serine, but rapid uptake was evident if sodium chloride was added to the assay mixture. Serine transport had an extremely poor affinity for sodium, and more than 30 mM was needed for half-maximal rates of uptake. Serine transport was inhibited by an excess of threonine, but an excess of alanine had little effect. Results indicated that S. bovis had separate sodium symport systems for serine or threonine and alanine, and either the membrane potential or chemical sodium gradient could drive uptake.
牛链球菌JB1细胞能够转运丝氨酸、苏氨酸或丙氨酸,但只有在钠缓冲液中孵育时才行。如果用磷酸钾洗涤经葡萄糖供能的细胞并悬浮于磷酸钾缓冲液中,则无法检测到摄取。用2-脱氧葡萄糖使细胞供能不足并在磷酸钠缓冲液中孵育的细胞仍能够转运丝氨酸,这一结果表明化学钠梯度能够驱动转运。然而,当供能不足的细胞用缬氨霉素处理并稀释到磷酸钠中以同时产生人工膜电位和化学钠梯度时,丝氨酸摄取速率比仅具有钠梯度的细胞高五倍。如果供能不足的细胞预先加载了钠(没有膜电位或钠梯度),则几乎没有丝氨酸转运。尼日利亚菌素和莫能菌素是能够逆转跨膜钠梯度的离子载体,它们强烈抑制三种氨基酸的钠依赖性摄取。装载了钾并稀释到氯化锂或氯化胆碱中的膜囊泡无法转运丝氨酸,但如果在测定混合物中加入氯化钠,则会有快速摄取。丝氨酸转运对钠的亲和力极低,半数最大摄取速率需要超过30 mM的钠。丝氨酸转运受到过量苏氨酸的抑制,但过量丙氨酸的影响很小。结果表明,牛链球菌具有用于丝氨酸或苏氨酸以及丙氨酸的独立钠同向转运系统,膜电位或化学钠梯度均可驱动摄取。