Probing Enzymatic Activity inside Living Cells Using a Nanowire鈥揅ell 鈥淪andwich鈥?Assay
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- 作者：Yu-Ran Na ; So Yeon Kim ; Jellert T. Gaublomme ; Alex K. Shalek ; Marsela Jorgolli ; Hongkun Park ; Eun Gyeong Yang
- 刊名：Nano Letters
- 出版年：2013
- 出版时间：January 9, 2013
- 年：2013
- 卷：13
- 期：1
- 页码：153-158
- 全文大小：305K
- 年卷期：v.13,no.1(January 9, 2013)
- ISSN：1530-6992
文摘
Developing a detailed understanding of enzyme function in the context of an intracellular signal transduction pathway requires minimally invasive methods for probing enzyme activity in situ. Here, we describe a new method for monitoring enzyme activity in living cells by sandwiching live cells between two vertical silicon nanowire (NW) arrays. Specifically, we use the first NW array to immobilize the cells and then present enzymatic substrates intracellularly via the second NW array by utilizing the NWs鈥?ability to penetrate cellular membranes without affecting cells鈥?viability or function. This strategy, when coupled with fluorescence microscopy and mass spectrometry, enables intracellular examination of protease, phosphatase, and protein kinase activities, demonstrating the assay鈥檚 potential in uncovering the physiological roles of various enzymes.