Jørgensen K E, Sheikh M I
Institute of Medical Biochemistry, University of Aarhus, Denmark.
Biochim Biophys Acta. 1988 Mar 3;938(3):345-52. doi: 10.1016/0005-2736(88)90132-0.
The characteristics of renal transport of pyruvate by luminal membrane vesicles from pars convoluta and pars recta of rabbit proximal tubule were studied. It was found that the uptake of pyruvate in these vesicle preparations occurred by means of multiple transport systems. An electrogenic and Na+-requiring system confined to pars convoluta, exists for transport of pyruvate with an intermediate affinity, KA = 0.71 +/- 0.08 mM. In vesicles from pars recta, the uptake of pyruvate was mediated by a dual transport system with a high (KA1 = 0.30 +/- 0.05 mM) and low affinity (KA2 = 5.75 +/- 0.82 mM). The relation of these three pyruvate transport systems to the transport of other monocarboxylates and dicarboxylates was determined by examination of the inhibitory effect of L-lactate, beta-hydroxybutyrate and L-malate on the renal uptake of pyruvate. It was found that L-malate efficiently reduced the uptake of pyruvate by vesicles from pars convoluta, while addition of monocarboxylates (L-lactate, beta-hydroxybutyrate) has no effect on the transport of pyruvate in this region of proximal tubule. Furthermore, it was observed that Na+-dependent uptake of L-malate was specifically inhibited to about the same extent by 1 mM pyruvate. These findings strongly suggest that pyruvate and L-malate are taken up by the same transport system in vesicles from pars convoluta, which is different from that of L-lactate and beta-hydroxybutyrate. Experiments designed to investigate the substrate specificity for the high and low affinity system for pyruvate in pars recta, revealed that monocarboxylates (L-lactate, beta-hydroxybutyrate) substantially inhibit pyruvate transport at the low, but not the high substrate concentration, indicating that all monocarboxylates compete for the same high-affinity transport system. By contrast, L-malate at low concentrations preferentially inhibited the low-affinity system for pyruvate. This observation led us to suggest that L-malate and pyruvate share a common transport system in pars recta with low affinity for pyruvate and high affinity for L-malate.
研究了兔近端小管曲部和直部的管腔膜囊泡对丙酮酸的肾转运特性。发现在这些囊泡制剂中,丙酮酸的摄取是通过多种转运系统进行的。一种局限于曲部的电生性且需要钠离子的系统,以中等亲和力(KA = 0.71 +/- 0.08 mM)存在,用于丙酮酸的转运。在直部的囊泡中,丙酮酸的摄取由一个双重转运系统介导,该系统具有高亲和力(KA1 = 0.30 +/- 0.05 mM)和低亲和力(KA2 = 5.75 +/- 0.82 mM)。通过检测L-乳酸、β-羟基丁酸和L-苹果酸对丙酮酸肾摄取的抑制作用,确定了这三种丙酮酸转运系统与其他单羧酸和二羧酸转运的关系。发现L-苹果酸能有效降低曲部囊泡对丙酮酸的摄取,而添加单羧酸(L-乳酸、β-羟基丁酸)对近端小管该区域的丙酮酸转运没有影响。此外,观察到1 mM丙酮酸能特异性地抑制L-苹果酸的钠离子依赖性摄取,且抑制程度大致相同。这些发现强烈表明,曲部囊泡中丙酮酸和L-苹果酸是通过同一转运系统摄取的,这与L-乳酸和β-羟基丁酸的转运系统不同。旨在研究直部丙酮酸高亲和力和低亲和力系统底物特异性的实验表明,单羧酸(L-乳酸、β-羟基丁酸)在低底物浓度下能显著抑制丙酮酸转运,但在高底物浓度下则无此作用,这表明所有单羧酸都竞争同一高亲和力转运系统。相比之下,低浓度的L-苹果酸优先抑制丙酮酸的低亲和力系统。这一观察结果使我们推测,在直部,L-苹果酸和丙酮酸共享一个对丙酮酸亲和力低而对L-苹果酸亲和力高的共同转运系统。