N-terminus regulation of VMAT2 mediates methamphetamine-stimulated efflux.
N-terminus regulation of VMAT2 mediates methamphetamine-stimulated efflux.
复制标题
VMAT2 的 N 端调节介导甲基苯丙胺刺激的外流。
DOI：
10.1016/j.neuroscience.2013.11.059
复制
发表时间：
2014
期刊：
影响因子：
3.3
通讯作者：
Ruoho,AE
中科院分区：
文献类型：
--
作者：
Torres,B;Ruoho,AE
研究了囊泡单胺转运蛋白2（VMAT 2）的N-末端20个氨基酸（AA）作为VMAT 2功能的调节剂。去除N-末端的前16或19个AA导致分子螯合[3 H]-5HT的能力降低。谷胱甘肽-S-转移酶的N-末端结构进行磷酸化的存在下，PKC在丝氨酸15和18。检查这些推定的磷酸化位点对功能的影响。在全长VMAT 2中用天冬氨酸对丝氨酸15和18进行磷酸模拟取代导致[3 H]-5HT螯合减少和甲基苯丙胺（METH）刺激的预载[3 H]-5HT流出减少。相反，丝氨酸15和18突变为丙氨酸保持了完整的净底物螯合，但消除了预积累的[3 H]-5HT的MET刺激外排。总之，这些数据表明了一种模型，其中VMAT 2 N-末端调节单胺螯合。
The 20 amino acid (AA) N-terminus of the vesicular monoamine transporter 2 (VMAT2) was examined as a regulator of VMAT2 function. Removal of the first 16 or 19 AAs of the N-terminus resulted in a molecule with reduced ability to sequester [3H]-5HT. A glutathione-S-transferase-construct of the N-terminus underwent phosphorylation in the presence of PKC at serines 15 and 18. These putative phosphorylation sites were examined for effects on function. Phospho-mimetic substitution of serines 15 and 18 with aspartate in the full-length VMAT2 resulted in reduced [3H]-5HT sequestration and reduced methamphetamine (METH)-stimulated efflux of preloaded [3H]-5HT. In contrast, mutation of serines 15 and 18 to alanines maintained intact net substrate sequestration but eliminated METH-stimulated efflux of pre-accumulated [3H]-5HT. In summary, these data suggest a model in which the VMAT2 N-terminus regulates monoamine sequestration.