Lo R S, Said H M, Unger T F, Hollander D, Miledi R
Medical Research Service, Veterans Administration Medical Center, Long Beach, California 90822.
Proc Biol Sci. 1991 Nov 22;246(1316):161-5. doi: 10.1098/rspb.1991.0139.
We investigated the existence of an endogenous uptake system for folate in Xenopus laevis oocytes. This was done by performing uptake measurements using [3H]folic acid. Uptake of folic acid was linear with time for 4 h of incubation, and was similar in collagenase-treated and non-treated oocytes. The uptake process was carrier-mediated, as suggested by the saturation of folic acid uptake with concentration, and by the ability of unlabelled folic acid and its related compounds to significantly inhibit the uptake of [3H]folic acid. The apparent Km and Vmax of the uptake process were 42 +/- 7 nM and 10.56 +/- 0.46 fmol per oocyte per 2 h, respectively. The uptake of folic acid was independent of the presence of Na+ in the incubation medium, but was highly pH dependent with severe inhibition occurring at pH lower than 6.5. Folic acid uptake was energy- and temperature-dependent, and was significantly inhibited by the anion transport inhibitors DIDS and SITS. These results demonstrate the existence of an endogenous carrier-mediated system for folic acid uptake in Xenopus oocytes. Further characterization of the molecular mechanism of folic acid uptake and its regulation in this non mammalian in vitro unicellular system may prove useful in furthering our understanding of folate movement across biological membranes.
我们研究了非洲爪蟾卵母细胞中叶酸内源性摄取系统的存在情况。这是通过使用[³H]叶酸进行摄取测量来完成的。叶酸摄取在孵育4小时内与时间呈线性关系,并且在胶原酶处理和未处理的卵母细胞中相似。正如叶酸摄取随浓度饱和以及未标记的叶酸及其相关化合物显著抑制[³H]叶酸摄取的能力所表明的那样,摄取过程是由载体介导的。摄取过程的表观Km和Vmax分别为42±7 nM和每2小时每个卵母细胞10.56±0.46 fmol。叶酸摄取与孵育培养基中Na⁺的存在无关,但高度依赖pH值,在pH低于6.5时会出现严重抑制。叶酸摄取依赖于能量和温度,并且被阴离子转运抑制剂DIDS和SITS显著抑制。这些结果证明了非洲爪蟾卵母细胞中存在一种内源性载体介导的叶酸摄取系统。在这个非哺乳动物体外单细胞系统中进一步表征叶酸摄取的分子机制及其调节,可能有助于增进我们对叶酸跨生物膜转运的理解。