Nord E P, Wright S H, Kippen I, Wright E M
J Membr Biol. 1983;72(3):213-21. doi: 10.1007/BF01870588.
The substrate specificity of a Na+-dependent transport pathway for L-lactate was studied in rabbit renal brush border membrane vesicles. Jmax for L-lactate transport was unaffected by the presence of a fixed concentration of two different short-chain monocarboxylic acids, while the apparent Kt(Ka) for L-lactate increased, and this is compatible with competitive inhibition. The inhibitor constants ("Ki"'s) for the transport pathway for the two solutes examined closely corresponded to the respective "Ki"'s derived from a Dixon plot. A broad range of compounds were then tested as potential inhibitors of L-lactate transport, and the "Ki"'s thereby derived yielded specific information regarding optimal substrate recognition by the carrier. A single carboxyl group is an absolute requirement for recognition, and preference is given to 3 to 6 C chain molecules. Addition of ketone, hydroxyl and, particularly, amine groups at any carbon position, diminishes substrate-carrier interaction. Intramolecular forces, notably the inductive effects of halogens, may play a role in enhancing substrate-carrier interaction; however, no correlation was found between pKa and "Ki" for the substrates examined. We conclude that a separate monocarboxylic acid transport pathway, discrete from either the D-glucose, alpha or beta neutral amino-acid, or dicarboxylic acid carriers, exists in the renal brush border, and this handles a broad range of monocarboxylates.
在兔肾刷状缘膜囊泡中研究了L-乳酸钠依赖性转运途径的底物特异性。L-乳酸转运的Jmax不受两种不同短链单羧酸固定浓度存在的影响,而L-乳酸的表观Kt(Ka)增加,这与竞争性抑制相符。所检测的两种溶质转运途径的抑制常数(“Ki”)与从Dixon图得出的各自“Ki”密切对应。然后测试了多种化合物作为L-乳酸转运的潜在抑制剂,由此得出的“Ki”提供了关于载体对最佳底物识别的具体信息。单个羧基是识别的绝对必要条件,并且优先选择3至6个碳链分子。在任何碳位置添加酮基、羟基,特别是氨基,都会减少底物与载体的相互作用。分子内作用力,特别是卤素的诱导效应,可能在增强底物与载体的相互作用中起作用;然而,在所检测的底物中,未发现pKa与“Ki”之间存在相关性。我们得出结论,肾刷状缘存在一条独立的单羧酸转运途径,与D-葡萄糖、α或β中性氨基酸或二羧酸载体不同,并且这条途径可处理多种单羧酸盐。