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

Nano Lett
Authors
Keywords
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.

Year of Publication
2013
Journal
Nano Lett
Volume
13
Issue
1
Pages
153-8
Date Published
2013 Jan 09
ISSN
1530-6992
DOI
10.1021/nl3037068
PubMed ID
23244056
PubMed Central ID
PMC3541459
Links
Grant list
5DP1OD003893-03 / OD / NIH HHS / United States
P50 HG006193 / HG / NHGRI NIH HHS / United States
DP1 DA035083 / DA / NIDA NIH HHS / United States
DP1 OD003893 / OD / NIH HHS / United States
1P50HG006193-01 / HG / NHGRI NIH HHS / United States