Probing enzymatic activity inside living cells using a nanowire-cell "sandwich" assay

Title
Probing enzymatic activity inside living cells using a nanowire-cell "sandwich" assay
Authors
나유란김소연Jellert T. GaublommeAlex K. ShalekMarsela JorgolliHongkun Park양은경
Keywords
Silicon nanowire; sandwich assembly; live cell assay; enzyme activity
Issue Date
2013-01
Publisher
Nano letters
Citation
VOL 13, NO 1, 153-158
Abstract
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.
URI
http://pubs.kist.re.kr/handle/201004/46282
ISSN
15306984
Appears in Collections:
KIST Publication > Article
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