Koomoa Dana-Lynn T, Musch Mark W, Goldstein Leon
Division of Biology and Medicine, Brown University, Providence, RI 02912, USA.
J Membr Biol. 2005 Dec;208(3):241-50. doi: 10.1007/s00232-005-7000-1. Epub 2006 Apr 7.
When swollen, skate red blood cells increase permeability and allow efflux of a number of solutes, including taurine. Hypoosmosis-induced taurine permeability appears to involve the red cell anion exchanger. However, three isoforms have been cloned from these cells. Therefore, to determine the ability of the individual isoform skate anion exchanger 1 (skAE1) to mediate hypoosmosis-induced taurine permeability as well as associated regulatory events, skAE1 was expressed in Xenopus oocytes. This study focused on investigating the role of tyrosine kinases and lipid rafts in the regulation of the channel. The results showed that tyrosine kinase inhibitors and lipid raft-disrupting agents inhibited the volume-sensitive organic osmolyte channel while protein tyrosine phosphatase inhibitors activated the channel in oocytes expressing skAE1. To study the role of lipid rafts in the activation of the volume-sensitive organic osmolyte channel, the cellular localization of skAE1 was investigated. Also, the role of tyrosine kinases was investigated by examining the tyrosine phosphorylation state of skAE1. Hypoosmotic stress induced mobilization of skAE1 into light membranes and the cell surface as well as tyrosine phosphorylation of skAE1. These events are involved in the activation of the volume-sensitive organic osmolyte channel in Xenopus oocytes expressing skAE1.
当肿胀时,鳐鱼红细胞会增加通透性,并允许包括牛磺酸在内的多种溶质流出。低渗诱导的牛磺酸通透性似乎涉及红细胞阴离子交换器。然而,已经从这些细胞中克隆出了三种同工型。因此,为了确定单个同工型鳐鱼阴离子交换器1(skAE1)介导低渗诱导的牛磺酸通透性以及相关调节事件的能力,skAE1在非洲爪蟾卵母细胞中进行了表达。本研究着重于研究酪氨酸激酶和脂筏在该通道调节中的作用。结果表明,酪氨酸激酶抑制剂和破坏脂筏的试剂抑制了体积敏感的有机渗透剂通道,而蛋白酪氨酸磷酸酶抑制剂则激活了表达skAE1的卵母细胞中的该通道。为了研究脂筏在激活体积敏感的有机渗透剂通道中的作用,对skAE1的细胞定位进行了研究。此外,通过检测skAE1的酪氨酸磷酸化状态来研究酪氨酸激酶的作用。低渗应激诱导skAE1向轻膜和细胞表面的移动以及skAE1的酪氨酸磷酸化。这些事件参与了表达skAE1的非洲爪蟾卵母细胞中体积敏感的有机渗透剂通道的激活。