Moseley R H, Takeda H, Zugger L J
Department of Internal Medicine, Veterans Affairs Medical Center, Ann Arbor, MI 48105, USA.
Hepatology. 1996 Jul;24(1):192-7. doi: 10.1002/hep.510240131.
Previous studies have demonstrated several pathways for the sinusoidal uptake of endogenous and exogenous organic cations, including two distinct organic cation:H+ exchanges and a separate carrier-mediated system driven by the inside-negative membrane potential. In this study, the driving force(s) for the uptake of the endogenous quaternary amine, choline, were determined in rat liver basolateral plasma membrane (blLPM) vesicles. Choline uptake into an osmotically sensitive space was not stimulated by an outwardly directed H+ or inwardly directed Na+ gradient. Instead, an inside-negative K+ diffusion potential stimulated choline uptake, suggesting the presence of a conductive pathway for choline uptake in bILPM vesicles. Conductive choline uptake was confirmed by inducing variable changes in the transmembrane potential with anions of different membrane permeability. Choline uptake in blLPM vesicles exhibitied 1) temperature dependency; 2) trans-stimulation; and 3) saturability, with an approximate Michaelis constant (Km) of 0.34 mmol/L and maximum velocity (Vmax) of 0.45 nmol/mg protein/15 s. Choline uptake in blLPM vesicles was cis-inhibited by the structurally similar derivative, hemicholinium-3 and acetylcholine, but not by substrates for other organic cation transport processes identified in blLPM vesicles, including thiamine, tetraethylammonium (TEA), tri-n-butyl-methylammonium (TBuMA), and N'-methylnicotinamide (NMN). These findings demonstrate an electrogenic pathway on the sinusoidal membrane for the uptake of this essential nutrient and support the existence of multiple pathways for the sinusoidal uptake of endogenous and exogenous organic cations.
先前的研究已经证明了内源性和外源性有机阳离子经窦状隙摄取的几种途径,包括两种不同的有机阳离子:H⁺交换以及由膜内负电位驱动的独立载体介导系统。在本研究中,测定了大鼠肝脏基底外侧质膜(blLPM)囊泡中内源性季铵盐胆碱摄取的驱动力。向渗透敏感空间内摄取胆碱不会受到外向H⁺梯度或内向Na⁺梯度的刺激。相反,膜内负的K⁺扩散电位刺激了胆碱摄取,这表明在bILPM囊泡中存在胆碱摄取的传导途径。通过用具有不同膜通透性的阴离子诱导跨膜电位的变化,证实了胆碱的传导性摄取。blLPM囊泡中的胆碱摄取表现出:1)温度依赖性;2)反式刺激;3)饱和性,其米氏常数(Km)约为0.34 mmol/L,最大速度(Vmax)为0.45 nmol/mg蛋白质/15秒。blLPM囊泡中的胆碱摄取受到结构相似的衍生物、半胱氨酸-3和乙酰胆碱的顺式抑制,但不受blLPM囊泡中鉴定出的其他有机阳离子转运过程的底物抑制,这些底物包括硫胺素、四乙铵(TEA)、三正丁基甲基铵(TBuMA)和N'-甲基烟酰胺(NMN)。这些发现证明了窦状隙膜上存在一种电生途径用于摄取这种必需营养素,并支持内源性和外源性有机阳离子经窦状隙摄取存在多种途径。