Wilde S W, Kilberg M S
Department of Biochemistry, J. Hillis Miller Health Center, University of Florida, Gainesville 32610.
Biochem J. 1991 Aug 1;277 ( Pt 3)(Pt 3):687-91. doi: 10.1042/bj2770687.
L-Glutamine, a major energy substrate for intestinal epithelial cells, can be extracted from intraluminal contents across the brush-border membrane and from arterial blood via the basolateral membrane. The purpose of the present study was to characterize glutamine transport by the basolateral membrane of rabbit epithelial cells. Transport of glutamine by isolated basolateral-membrane vesicles was mediated by both Na(+)-dependent and Na(+)-independent carriers. Tests were performed to distinguish glutamine uptake by likely transport agencies, including Systems A, ASC, N, IMINO, NBB, L and asc. The Na(+)-dependent glutamine uptake was strongly inhibited by an excess of 2-(methylamino)isobutyric acid (MeAIB), and glutamine was equally effective in inhibiting MeAIB transport. The reciprocal inhibition analysis, as well as a sensitivity to increased H+ concentration, indicates that Na(+)-dependent glutamine transport across the basolateral membrane is mediated by System A. The saturable Na(+)-independent glutamine transport was markedly inhibited by 2-aminobicyclo-[2,2,1]-heptane-2-carboxylic acid ('BCH') and insensitive to changes in assay pH, suggesting uptake via System L rather than System asc. The presence of a Na(+)-dependent carrier to mediate active transport of glutamine across the basolateral membrane is probably essential to ensure a continuous supply of this vital substrate to the enterocyte in the post-absorptive state.
L-谷氨酰胺是肠上皮细胞的主要能量底物,可通过刷状缘膜从肠腔内内容物中摄取,并通过基底外侧膜从动脉血中摄取。本研究的目的是表征兔上皮细胞基底外侧膜对谷氨酰胺的转运。分离的基底外侧膜囊泡对谷氨酰胺的转运由钠依赖性和非钠依赖性载体介导。进行了测试以区分可能的转运机制对谷氨酰胺的摄取,包括系统A、ASC、N、亚氨基、NBB、L和asc。过量的2-(甲氨基)异丁酸(MeAIB)强烈抑制钠依赖性谷氨酰胺摄取,谷氨酰胺在抑制MeAIB转运方面同样有效。相互抑制分析以及对增加的H⁺浓度的敏感性表明,钠依赖性谷氨酰胺跨基底外侧膜的转运由系统A介导。可饱和的非钠依赖性谷氨酰胺转运受到2-氨基双环-[2,2,1]-庚烷-2-羧酸(“BCH”)的显著抑制,并且对测定pH的变化不敏感,表明是通过系统L而非系统asc进行摄取。存在一种钠依赖性载体来介导谷氨酰胺跨基底外侧膜的主动转运,这可能对于确保在吸收后状态下向肠细胞持续供应这种重要底物至关重要。