Kragh-Hansen U, Sheikh M I
J Physiol. 1984 Sep;354:55-67. doi: 10.1113/jphysiol.1984.sp015361.
The mechanism of renal transport of L- and D-serine by membrane vesicles prepared from either whole cortex, pars convoluta or pars recta of rabbit proximal tubule was studied by a rapid filtration technique and by a spectrophotometric method using a potential-sensitive carbocyanine dye. Transport studies carried out with different salt gradients and by employing various ionophores showed that uptake of both L- and D-serine by luminal membrane vesicles from whole cortex was mediated by an Na+-dependent and electrogenic transport process. Eadie-Hofstee analysis of experimental data, obtained under extravesicular greater than intravesicular NaCl gradients, revealed the existence of multiple transport systems for L-serine but only one system for the D-isomer. The value of KA (the concentration producing a half-maximal optical response) for the D-serine transport system was calculated to be approximately 30 mM. Luminal membrane vesicles from pars convoluta take up both L- and D-serine by a single and common transport system. KA values for L- and D-serine transport were calculated to be 3.7 and 30 mM, respectively. Luminal membrane vesicles from pars recta take up L-serine by means of two transport systems, one of high affinity (KA = 0.37 mM) and the other of low affinity (KA = 10 mM). By contrast, no D-serine transport by these membrane vesicles could be detected. Uptake of L-serine by basolateral membrane vesicles is Na+ independent and electroneutral. Filtration studies showed that the transport is saturable (Km = 25-30 mM) and is inhibited by the presence of L-phenylalanine (but not by D-serine), indicating carrier-mediated uptake of L-serine.
采用快速过滤技术和使用电位敏感碳氰染料的分光光度法,研究了从兔近端小管全皮质、曲部或直部制备的膜囊泡对L-丝氨酸和D-丝氨酸的肾转运机制。在不同盐梯度下并使用各种离子载体进行的转运研究表明,全皮质腔面膜囊泡对L-丝氨酸和D-丝氨酸的摄取是由Na⁺依赖性和电生转运过程介导的。在囊泡外NaCl梯度大于囊泡内的情况下获得的实验数据的伊迪-霍夫斯泰分析表明,L-丝氨酸存在多个转运系统,而D-异构体只有一个系统。计算出D-丝氨酸转运系统的KA值(产生最大光学响应一半时的浓度)约为30 mM。曲部的腔面膜囊泡通过单一的共同转运系统摄取L-丝氨酸和D-丝氨酸。计算出L-丝氨酸和D-丝氨酸转运的KA值分别为3.7 mM和30 mM。直部的腔面膜囊泡通过两种转运系统摄取L-丝氨酸,一种是高亲和力的(KA = 0.37 mM),另一种是低亲和力的(KA = 10 mM)。相比之下,未检测到这些膜囊泡对D-丝氨酸的转运。基底外侧膜囊泡对L-丝氨酸的摄取不依赖于Na⁺且是电中性的。过滤研究表明,转运是可饱和的(Km = 25 - 30 mM),并且受到L-苯丙氨酸(而不是D-丝氨酸)的抑制,表明L-丝氨酸是通过载体介导摄取的。