Probing enzymatic activity inside living cells using a nanowire-cell "sandwich" assay
- Probing enzymatic activity inside living cells using a nanowire-cell "sandwich" assay
- 나유란; 김소연; Jellert T. Gaublomme; Alex K. Shalek; Marsela Jorgolli; Hongkun Park; 양은경
- Silicon nanowire; sandwich assembly; live cell assay; enzyme activity
- Issue Date
- Nano letters
- VOL 13, NO 1, 153-158
- 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’ ability to penetrate
cellular membranes without affecting cells’ 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’s potential in uncovering the physiological roles of various enzymes.
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