Said H M, Redha R
Department of Pediatric Gastroenterology, Vanderbilt University School of Medicine, Nashville, TN 37232.
Biochem J. 1987 Oct 1;247(1):141-6. doi: 10.1042/bj2470141.
The mechanism of exit of folate from the enterocyte, i.e. transport across the basolateral membrane, is not known. In this study we examined, using basolateral membrane vesicles, the transport of folic acid across the basolateral membrane of rat intestine. Uptake of folic acid by these vesicles represents transport of the substrate into the intravesicular compartment and not binding to the membrane surface. The rate of folic acid transport was linear for the first 1 min of incubation but decreased thereafter, reaching equilibrium after 5 min of incubation. The transport of folic acid was: (1) saturable as a function of concentration with an apparent Km of 0.6 +/- 0.17 microM and Vmax. of 1.01 +/- 0.11 pmol/30 s per mg of protein; (2) inhibited in a competitive manner by the structural analogues 5-methyltetrahydrofolate and methotrexate (Ki = 2 and 1.4 microM, respectively); (4) electroneutral; (5) Na+-independent; (6) sensitive to the effect of the anion exchange inhibitor 4,4'-di-isothiocyanatostilbene-2,2'-disulphonic acid (DIDS). These data indicate the existence of a carrier-mediated transport system for folic acid in rat intestinal basolateral membrane and demonstrate that the transport process is electroneutral, Na+-independent and sensitive to the effect of anion exchange inhibition.
叶酸从肠细胞排出的机制,即跨越基底外侧膜的转运,尚不清楚。在本研究中,我们使用基底外侧膜囊泡研究了叶酸在大鼠肠道基底外侧膜的转运。这些囊泡对叶酸的摄取代表底物转运至囊泡内区室,而非与膜表面结合。叶酸转运速率在孵育的最初1分钟呈线性,但此后下降,孵育5分钟后达到平衡。叶酸的转运具有以下特点:(1)作为浓度的函数呈饱和状态,表观Km为0.6±0.17微摩尔,Vmax为每毫克蛋白质1.01±0.11皮摩尔/30秒;(2)被结构类似物5-甲基四氢叶酸和甲氨蝶呤以竞争性方式抑制(Ki分别为2和1.4微摩尔);(4)电中性;(5)不依赖钠离子;(6)对阴离子交换抑制剂4,4'-二异硫氰酸根合芪-2,2'-二磺酸(DIDS)的作用敏感。这些数据表明大鼠肠道基底外侧膜存在一种载体介导的叶酸转运系统,并证明该转运过程是电中性的、不依赖钠离子的,且对阴离子交换抑制作用敏感。