Anterograde and retrograde intracellular trafficking of fluorescent cellular prion protein
Anterograde and retrograde intracellular trafficking of fluorescent cellular prion protein
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DOI：
10.1016/j.bbrc.2004.01.126
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发表时间：
2004-03-19
影响因子：
3.1
通讯作者：
Kaneko, K
中科院分区：
文献类型：
--
作者：
Hachiya, NS;Watanabe, K;Kaneko, K
为了研究NH 2-末端细胞朊病毒蛋白（PrPC）片段的微管相关细胞内运输[Biochem.Biophys.2004]，通信资源313（2004）818]，我们在活细胞中进行荧光PrPC（GFP-PrPC）的实时成像。这种GFP-PrPC以140180 nm/s的速度向质膜方向顺行运动，并以1.0-1.2 μ m/s的速度向内逆行运动。GFP-PrPC的顺向和逆向运动分别被驱动蛋白家族抑制剂（AMP-PNP）和动力蛋白家族抑制剂（钒酸盐）阻断。此外，抗驱动蛋白抗体（α-驱动蛋白）阻断其顺行运动，而抗动力蛋白抗体（α-动力蛋白）阻断其逆行运动。这些数据表明GFP-PrPC的驱动蛋白家族驱动的顺行和动力蛋白驱动的逆行运动。PrPC的相互作用结构域的作图鉴定了与驱动蛋白家族相互作用所必需的氨基酸残基：NH 2-末端小鼠（Mo）残基53-91和动力蛋白：NH 2-末端Mo残基23-33。我们的研究结果认为，离散的N-末端氨基酸残基是必不可少的顺行和逆行的细胞内运动的PrPC。(C)2004年爱思唯尔公司All rights reserved.
In order to investigate the microtubule-associated intracellular trafficking of the NH2-terminal cellular prion protein (PrPC) fragment [Biochem. Biophys. Res. Commun. 313 (2004) 818], we performed a real-time imaging of fluorescent PrPC (GFP-PrPC) in living cells. Such GFP-PrPC exhibited an anterograde movement towards the direction of plasma membranes at a speed of 140180 nm/s, and a retrograde movement inwardly at a speed of 1.0-1.2 mum,/s. The anterograde and retrograde movements of GFP-PrPC were blocked by a kinesin family inhibitor (AMP-PNP) and a dynein family inhibitor (vanadate), respectively. Furthermore, anti-kinesin antibody (alpha-kinesin) blocked its anterograde motility, whereas anti-dynein antibody (alpha-dynein) blocked its retrograde motility. These data suggested the kinesin family-driven anterograde and the dynein-driven retrograde movements of GFP-PrPC. Mapping of the interacting domains of PrPC identified amino acid residues indispensable for interactions with kinesin family: NH2-terminal mouse (Mo) residues 53-91 and dynein: NH2-terminal Mo residues 23-33, respectively. Our findings argue that the discrete N-terminal amino acid residues are indispensable for the anterograde and retrograde intracellular movements of PrPC. (C) 2004 Elsevier Inc. All rights reserved.