Ono Junichiro, Fushimi Satoko, Suzuki Shingo, Ameno Kiyoshi, Kinoshita Hiroshi, Shirakami Gotaro, Kabayama Kazuya
Department of Anesthesiology Faculty of Medicine Kagawa University Kita-gun Kagawa Japan.
Department of Anesthesiology KKR Takamatsu Hospital Kagawa Japan.
FEBS Open Bio. 2018 May 23;8(7):1127-1134. doi: 10.1002/2211-5463.12443. eCollection 2018 Jul.
The volatile anesthetic isoflurane (ISO) has previously been shown to increase the fluidity of artificial lipid membranes, but very few studies have used biological cell membranes. Therefore, to investigate whether ISO affects the mobility of membrane proteins, fluorescence-labeled transferrin receptor (TfR) and glycosylphosphatidylinositol (GPI)-anchored protein were expressed in human embryonic kidney 293T cells and neural cells and lateral diffusion was examined using fluorescence recovery after photobleaching. Lateral diffusion of the TfR increased with ISO treatment. On the other hand, there was no effect on GPI-anchored protein. We also used GC/MS to confirm that there was no change in the concentration of ISO due to vaporization during measurement. These results suggest that ISO affects the mobility of transmembrane protein molecules in living cells.
挥发性麻醉剂异氟烷(ISO)此前已被证明可增加人工脂质膜的流动性,但很少有研究使用生物细胞膜。因此,为了研究ISO是否影响膜蛋白的流动性,在人胚肾293T细胞和神经细胞中表达了荧光标记的转铁蛋白受体(TfR)和糖基磷脂酰肌醇(GPI)锚定蛋白,并使用光漂白后的荧光恢复来检测横向扩散。经ISO处理后,TfR的横向扩散增加。另一方面,对GPI锚定蛋白没有影响。我们还使用气相色谱/质谱法确认测量过程中由于蒸发导致ISO浓度没有变化。这些结果表明,ISO会影响活细胞中跨膜蛋白分子的流动性。