Maenz D D, Chenu C, Breton S, Berteloot A
Department of Physiology, Faculty of Medicine, Université de Montréal, Québec, Canada.
J Biol Chem. 1992 Jan 25;267(3):1510-6.
Initial rates of Na(+)-dependent L-glutamic and D-aspartic acid uptake were determined at various substrate concentrations using a fast sampling, rapid filtration apparatus, and the resulting data were analyzed by nonlinear computer fitting to various transport models. At pH 6.0, L-glutamic acid transport was best accounted for by the presence of both high (Km = 61 microM) and low (Km = 7.0 mM) affinity pathways, whereas D-aspartic acid transport was restricted to a single high affinity route (Km = 80 microM). Excess D-aspartic acid and L-phenylalanine served to isolate L-glutamic acid flux through the remaining low and high affinity systems, respectively. Inhibition studies of other amino acids and analogs allowed us to identify the high affinity pathway as the X-AG system and the low affinity one as the intestinal NBB system. The pH dependences of the high and low affinity pathways of L-glutamic acid transport also allowed us to establish some relationship between the NBB and the more classical ASC system. Finally, these studies also revealed a heterotropic activation of the intestinal X-AG transport system by all neutral amino acids but glycine through an apparent activation of Vmax.
使用快速取样、快速过滤装置在不同底物浓度下测定了Na⁺依赖性L-谷氨酸和D-天冬氨酸摄取的初始速率,并通过非线性计算机拟合各种转运模型对所得数据进行分析。在pH 6.0时,L-谷氨酸转运最好由高亲和力途径(Km = 61 μM)和低亲和力途径(Km = 7.0 mM)共同解释,而D-天冬氨酸转运则局限于单一的高亲和力途径(Km = 80 μM)。过量的D-天冬氨酸和L-苯丙氨酸分别用于分离L-谷氨酸通过其余低亲和力和高亲和力系统的通量。对其他氨基酸及其类似物的抑制研究使我们能够将高亲和力途径鉴定为X-AG系统,将低亲和力途径鉴定为肠道NBB系统。L-谷氨酸转运的高亲和力和低亲和力途径对pH的依赖性也使我们能够确定NBB与更经典的ASC系统之间的一些关系。最后,这些研究还揭示了除甘氨酸外的所有中性氨基酸通过明显激活Vmax对肠道X-AG转运系统的异促激活作用。