Transient Photoinactivation of Cell Membrane Protein Activity without Genetic Modification by Molecular Hyperthermia
Transient Photoinactivation of Cell Membrane Protein Activity without Genetic Modification by Molecular Hyperthermia
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DOI：
10.1021/acsnano.9b01993
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发表时间：
2019-11-01
期刊：
影响因子：
17.1
通讯作者：
Qin, Zhenpeng
中科院分区：
文献类型：
--
作者：
Kang, Peiyuan;Li, Xiaoqing;Qin, Zhenpeng
在生物系统中对蛋白质活性进行精确调控在生物医学科学中具有广泛应用。然而，在不进行基因改造的情况下利用光来调控生物系统中的蛋白质活性具有挑战性。在此，我们报道了一种以高时空分辨率在活细胞中光学关闭蛋白质活性的技术，称为分子热疗（MH）。MH基于通过短激光脉冲对等离子体金纳米粒子进行纳米级限域加热，使目标蛋白质解折叠并光失活。首先，我们表明蛋白酶激活受体2（PAR2），一种G蛋白偶联受体以及导致痛觉敏化的重要途径，可以通过MH在原位光失活，且不影响细胞增殖。可以在激光靶向的细胞中关闭PAR2活性，而不影响周围细胞。此外，我们通过使PAR2失活而保持其他受体完整，证明了MH的分子特异性。其次，我们证明了脑内皮单层中紧密连接蛋白的光失活会导致体外血脑屏障可逆性开放。最后，MH导致的蛋白质失活低于纳米气泡产生阈值，因此主要是由于纳米级加热。MH在时间和长度尺度上都不同于传统热疗（传统热疗会引起整体组织加热）：纳秒对秒，纳米对毫米。我们的结果表明，MH能够在不进行基因改造的情况下对蛋白质活性和细胞行为进行选择性和远程调控。
Precise manipulation of protein activity in living systems has broad applications in biomedical sciences. However, it is challenging to use light to manipulate protein activity in living systems without genetic modification. Here, we report a technique to optically switch off protein activity in living cells with high spatiotemporal resolution, referred to as molecular hyperthermia (MH). MH is based on the nanoscale-confined heating of plasmonic gold nanoparticles by short laser pulses to unfold and photoinactivate targeted proteins of interest. First, we show that protease-activated receptor 2 (PAR2), a G-protein-coupled receptor and an important pathway that leads to pain sensitization, can be photoinactivated in situ by MH without compromising cell proliferation. PAR2 activity can be switched off in laser-targeted cells without affecting surrounding cells. Furthermore, we demonstrate the molecular specificity of MH by inactivating PAR2 while leaving other receptors intact. Second, we demonstrate that the photoinactivation of a tight junction protein in brain endothelial monolayers leads to a reversible blood-brain barrier opening in vitro. Lastly, the protein inactivation by MH is below the nanobubble generation threshold and thus is predominantly due to the nanoscale heating. MH is distinct from traditional hyperthermia (that induces global tissue heating) in both its time and length scales: nanoseconds versus seconds, nanometers versus millimeters. Our results demonstrate that MH enables selective and remote manipulation of protein activity and cellular behavior without genetic modification.