Fan M Z, Adeola O, McBurney M I, Cheeseman C I
Department of Animal Sciences, Purdue University, West Lafayette, IN 47907, USA.
Comp Biochem Physiol A Mol Integr Physiol. 1998 Dec;121(4):411-22. doi: 10.1016/s1095-6433(98)10152-6.
L-Glutamine transport into porcine jejunal enterocyte brush border membrane vesicles was studied. Uptake was mediated by a Na(+)-dependent and a Na(+)-independent pathway as well as by diffusion. The initial rates of glutamine uptake over a range of concentrations is both Na(+)-gradient and Na(+)-free conditions were analyzed and kinetic parameters were obtained. Na(+)-dependent glutamine transport had a K(m) of 0.77 +/- 0.16 mM and a Jmax of 70.7 +/- 5.8 pmol mg protein-1 s-1; Na(+)-independent glutamine transport had a K(m) of 3.55 +/- 0.78 mM and a Jmax of 55.1 +/- 6.6 pmol mg protein-1 s-1. The non-saturable component measured with HgCl2-poisoned brush border membrane vesicles in the Na(+)-free condition contained passive diffusion and non-specific membrane binding and was defined to be apparent glutamine diffusion and the glutamine permeability coefficient (Kdiff) was estimated to be Kdiff = 3.78 +/- 0.06 pmol 1 mg protein-1 mmol-1 s-1. Results of inhibition experiments showed that Na(+)-dependent glutamine uptake occurred primarily through the brush border system-B degree transporters, whereas Na(+)-independent glutamine uptake occurred via the system-L transporters. Furthermore, the kinetics of L-leucine and L-cysteine inhibition of L-glutamine uptake demonstrated that neutral amino acids sharing the same brush border transporters can effectively inhibit each other in their transport.
研究了L-谷氨酰胺转运至猪空肠肠上皮细胞刷状缘膜囊泡的过程。摄取由Na⁺依赖性和Na⁺非依赖性途径以及扩散介导。分析了在一系列浓度下谷氨酰胺摄取在有Na⁺梯度和无Na⁺条件下的初始速率,并获得了动力学参数。Na⁺依赖性谷氨酰胺转运的K(m)为0.77±0.16 mM,Jmax为70.7±5.8 pmol mg蛋白⁻¹ s⁻¹;Na⁺非依赖性谷氨酰胺转运的K(m)为3.55±0.78 mM,Jmax为55.1±6.6 pmol mg蛋白⁻¹ s⁻¹。在无Na⁺条件下用HgCl₂中毒的刷状缘膜囊泡测量的非饱和成分包含被动扩散和非特异性膜结合,被定义为表观谷氨酰胺扩散,谷氨酰胺渗透系数(Kdiff)估计为Kdiff = 3.78±0.06 pmol 1 mg蛋白⁻¹ mmol⁻¹ s⁻¹。抑制实验结果表明,Na⁺依赖性谷氨酰胺摄取主要通过刷状缘系统-B°转运体发生,而Na⁺非依赖性谷氨酰胺摄取通过系统-L转运体发生。此外,L-亮氨酸和L-半胱氨酸对L-谷氨酰胺摄取的抑制动力学表明,共用相同刷状缘转运体的中性氨基酸在转运过程中可有效相互抑制。