Gilles Wolfgang, Vulcu Sebastian D, Liewald Jana F, Habermeier Alice, Vékony Nicole, Closs Ellen I, Rupp Johanna, Nawrath Hermann
Department of Pharmacology, Johannes-Gutenberg University, Obere Zahlbacher Str. 67, 55101 Mainz, Germany.
Biochim Biophys Acta. 2005 Mar 1;1668(2):234-9. doi: 10.1016/j.bbamem.2004.12.011. Epub 2005 Jan 21.
hCAT-3 (human cationic amino acid transporter type three) was investigated with both the two-electrode voltage clamp method and tracer experiments. Oocytes expressing hCAT-3 displayed less negative membrane potentials and larger voltage-dependent currents than native or water-injected oocytes did. Ion substitution experiments in hCAT-3-expressing oocytes revealed a large conductance for Na+ and K+. In the presence of L-Arg, voltage-dependent inward and outward currents were observed. At symmetrical (inside/outside) concentrations of L-Arg, the conductance of the transporter increased monoexponentially with the L-Arg concentrations; the calculated Vmax and KM values amounted to 8.3 microS and 0.36 mM, respectively. The time constants of influx and efflux of [3H]L-Arg, at symmetrically inside/outside L-Arg concentrations (1 mM), amounted to 79 and 77 min, respectively. The flux data and electrophysiological experiments suggest that the transport of L-Arg through hCAT-3 is symmetric, when the steady state of L-Arg flux has been reached. It is concluded that hCAT-3 is a passive transport system that conducts monovalent cations including L-Arg. The particular role of hCAT-3 in the diverse tissues remains to be elucidated.
采用双电极电压钳法和示踪实验对hCAT-3(人类阳离子氨基酸转运体3型)进行了研究。与天然或注射水的卵母细胞相比,表达hCAT-3的卵母细胞表现出较小的负膜电位和较大的电压依赖性电流。在表达hCAT-3的卵母细胞中进行的离子替代实验显示,其对Na+和K+具有较大的电导。在L-精氨酸存在的情况下,观察到了电压依赖性内向和外向电流。在L-精氨酸的对称(膜内/膜外)浓度下,转运体的电导随L-精氨酸浓度呈单指数增加;计算得出的Vmax和KM值分别为8.3微西门子和0.36毫摩尔。在对称的膜内/膜外L-精氨酸浓度(1毫摩尔)下,[3H]L-精氨酸流入和流出的时间常数分别为79分钟和77分钟。通量数据和电生理实验表明,当达到L-精氨酸通量的稳态时,L-精氨酸通过hCAT-3的转运是对称的。结论是,hCAT-3是一个传导包括L-精氨酸在内的单价阳离子的被动转运系统。hCAT-3在不同组织中的具体作用仍有待阐明。