Sopanen T, Väisänen E
Biotechnical Laboratory, Technical Research Centre of Finland, Tietotie 2, SF-02150 Espoo 15, Finland.
Plant Physiol. 1985 Aug;78(4):684-9. doi: 10.1104/pp.78.4.684.
Scutella separated from germinating grains of barley (Hordeum vulgare L. cv Himalaya) took up [(14)C]glutamine at an initial rate of about 10 micromoles.gram(-1).hour(-1) in the standard assay conditions (pH 5, 30 degrees C, 1 millimolar glutamine). Inhibition by unlabeled glutamine and by dinitrophenol indicated that about 95% of the uptake was due to carrier-mediated active transport. The pH optimum of the uptake was 5, and after correction for a nonmediated component the uptake appeared to conform to Michaelis-Menten kinetics with an apparent K(m) of about 2 millimolar and a V(max) of about 25 micromoles.gram(-1).hour(-1).The uptake of glutamine was inhibited by all of the 18 amino acids tested; the mode of inhibition was studied only with proline and was competitive. Eight of the ten amino acids tested at high concentrations appeared to be able to inhibit the mediated uptake of glutamine virtually completely. However, when the inhibitory effect of asparagine was extrapolated to an infinitely high concentration of asparagine, about 24% of the mediated uptake of glutamine remained uninhibited. These results suggest that glutamine is taken up by two (or more) rather unspecific amino acid uptake systems, the minor one having no affinity for asparagine.Glutamine and alanine could completely inhibit the mediated uptake of 1 millimolar leucine, but about 12% of the mediated uptake appeared to be uninhibitable by asparagine. Furthermore, the ratio of the mediated uptake of glutamine to that of leucine changed from 0.9 to 1.7 between days 1 and 3 of germination. These results give further support for the presence of two unspecific amino acid uptake systems in barley scutella.
从发芽的大麦(Hordeum vulgare L. cv Himalaya)种子中分离出的盾片,在标准测定条件下(pH 5、30℃、1毫摩尔谷氨酰胺),以约10微摩尔·克⁻¹·小时⁻¹的初始速率摄取[(¹⁴)C]谷氨酰胺。未标记的谷氨酰胺和二硝基苯酚的抑制作用表明,约95%的摄取是由载体介导的主动转运引起的。摄取的最适pH为5,校正非介导成分后,摄取似乎符合米氏动力学,表观K(m)约为2毫摩尔,V(max)约为25微摩尔·克⁻¹·小时⁻¹。所测试的18种氨基酸均抑制谷氨酰胺的摄取;仅对脯氨酸研究了抑制模式,为竞争性抑制。在高浓度下测试的10种氨基酸中有8种似乎能够几乎完全抑制谷氨酰胺的介导摄取。然而,当将天冬酰胺的抑制作用外推到无限高浓度的天冬酰胺时,约24%的谷氨酰胺介导摄取仍未被抑制。这些结果表明,谷氨酰胺是通过两个(或更多)相当非特异性的氨基酸摄取系统摄取的,较小的那个系统对天冬酰胺没有亲和力。谷氨酰胺和丙氨酸可以完全抑制1毫摩尔亮氨酸的介导摄取,但约12%的介导摄取似乎不受天冬酰胺抑制。此外,在发芽的第1天到第3天之间,谷氨酰胺介导摄取与亮氨酸介导摄取的比率从0.9变为1.7。这些结果进一步支持了大麦盾片中存在两个非特异性氨基酸摄取系统。