Iioka H, Moriyama I, Saito M, Hino K, Okamura Y, Ichijo M
Nihon Sanka Fujinka Gakkai Zasshi. 1986 Apr;38(4):529-34.
Using microvillous membrane vesicles prepared from human normal early and full term placenta, we investigated the transport mechanism of L-alanine and the change in its transport activity during gestation. We estimated the purity of microvillous membrane vesicles prepared from human placenta from the relative specific activities (microvilli versus homogenate) of the membrane's maker enzymes, alkaline phosphatase (ALP), 5'-nucleotidase, and gamma-glutamyltranspeptidase ( gamma-GTP). In early pregnancy (12-15 weeks gestational age), the relative specific activities (microvilli versus homogenate) were calculated to be: ALP: 15.3, 5'-nucleotidase: 14.0, gamma-GTP: 8.3, and in full term pregnancy (37-40 weeks gestational age) the relative specific activities (microvilli versus homogenate) were calculated to be: ALP: 16.0, 5'-nucleotidase: 14.8, gamma-GTP: 7.5. The uptake of L-alanine into microvillous membrane vesicles was Na+ electrochemical gradient (extravesicular greater than intravesicular) dependent and this Na+ dependent uptake was membrane-potentially sensitive both in early pregnancy and in full term pregnancy. The kinetics parameter of the initial L-alanine uptake into microvillous membrane vesicles were calculated to be: Km: 0.78 +/- 0.20 mM, Vmax: 0.62 +/- 0.21 nmol/mg protein/20 sec in early pregnancy, Km: 0.80 +/- 0.24 mM, Vmax: 3.53 +/- 0.70 nmol/mg protein/20 sec in full term pregnancy. In conclusion, the placental transport mechanisms of L-alanine in both early and full term pregnancy were the same, and the L-alanine transport activity of full term pregnancy was much greater than that of early pregnancy.
我们使用从人类正常早期和足月胎盘制备的微绒毛膜囊泡,研究了L-丙氨酸的转运机制及其在妊娠期间转运活性的变化。我们根据膜标记酶碱性磷酸酶(ALP)、5'-核苷酸酶和γ-谷氨酰转肽酶(γ-GTP)的相对比活性(微绒毛与匀浆)估算了从人胎盘制备的微绒毛膜囊泡的纯度。在妊娠早期(妊娠12 - 15周),计算得到的相对比活性(微绒毛与匀浆)为:ALP:15.3,5'-核苷酸酶:14.0,γ-GTP:8.3;在足月妊娠(妊娠37 - 40周),计算得到的相对比活性(微绒毛与匀浆)为:ALP:16.0,5'-核苷酸酶:14.8,γ-GTP:7.5。L-丙氨酸进入微绒毛膜囊泡的摄取依赖于Na⁺电化学梯度(囊泡外大于囊泡内),并且这种Na⁺依赖性摄取在妊娠早期和足月妊娠时均对膜电位敏感。计算得到L-丙氨酸初始摄取进入微绒毛膜囊泡的动力学参数为:妊娠早期,Km:0.78±0.20 mM,Vmax:0.62±0.21 nmol/mg蛋白/20秒;足月妊娠时,Km:0.80±0.24 mM,Vmax:3.53±0.70 nmol/mg蛋白/20秒。总之,妊娠早期和足月妊娠时胎盘对L-丙氨酸的转运机制相同,且足月妊娠时L-丙氨酸的转运活性远高于妊娠早期。