Borgese Franck, Renard Céline, Gabillat Nicole, Pellissier Bernard, Guizouarn Hélène
Laboratoire de Physiologie des Membranes Cellulaires, FRE 2721, CNRS-Université de Nice, Bâtiment Jean Maetz, 284 chemin du Lazaret, 06230 Villefranche/Mer, France.
Biochim Biophys Acta. 2004 Jul 1;1664(1):80-7. doi: 10.1016/j.bbamem.2004.04.007.
It was previously shown that expressed in Xenopus oocyte the trout (tAE1) and the mouse (mAE1) anion exchangers behave differently: both elicit anion exchange activity but only tAE1 induces a transport of organic solutes correlated with an anion conductance. In order to identify the structural domains involved in the induction of tAE1 channel activity, chimeras have been prepared between mouse and trout AE1. As some constructs were not expressed at the plasma membrane, skate exchanger (skAE1) was used instead of mouse exchanger to complete the structure-function analysis. The present paper shows that skAE1, highly similar to mAE1, does not induce a chloride conductance when expressed in Xenopus oocyte. Construct expression analysis showed that only tAE1 transmembrane domain is linked to the anion conductance. More precisely, we identified two regions composed of helices 6, 7 and 8 and putative helices 12 and 13 which are required for this function.
先前的研究表明,在非洲爪蟾卵母细胞中表达的鳟鱼(tAE1)和小鼠(mAE1)阴离子交换蛋白表现不同:二者均引发阴离子交换活性,但只有tAE1能诱导与阴离子电导相关的有机溶质转运。为了确定参与诱导tAE1通道活性的结构域,制备了小鼠和鳟鱼AE1之间的嵌合体。由于一些构建体未在质膜上表达,因此使用鳐鱼交换蛋白(skAE1)代替小鼠交换蛋白来完成结构-功能分析。本文表明,与mAE1高度相似的skAE1在非洲爪蟾卵母细胞中表达时不会诱导氯离子电导。构建体表达分析表明,只有tAE1跨膜结构域与阴离子电导相关。更确切地说,我们确定了由螺旋6、7和8以及假定的螺旋12和13组成的两个区域,这两个区域是该功能所必需的。