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Methods Enzymol. 2014;544:271-97. doi: 10.1016/B978-0-12-417158-9.00011-X.

Single-molecule sensing of caspase activation in live cells via plasmon coupling nanotechnology.

Author information

1
Department of Pharmaceutical Chemistry, University of California, San Francisco, California, USA; Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, California, USA.
2
Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, California, USA; Department of Otolaryngology, University of California, San Francisco, California, USA.
3
Department of Pharmaceutical Chemistry, University of California, San Francisco, California, USA; Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, California, USA. Electronic address: charles.craik@ucsf.edu.

Abstract

Apoptotic caspases execute programmed cell death, where low levels of caspase activity are linked to cancer (Kasibhatla & Tseng, 2003). Chemotherapies utilize induction of apoptosis as a key mechanism for cancer treatment, where caspase-3 is a major player involved in dismantling these aberrant cells. The ability to sensitively measure the initial caspase-3 cleavage events during apoptosis is important for understanding the initiation of this complex cellular process; however, current ensemble methods are not sensitive enough to measure single cleavage events in cells. To overcome this, we describe a procedure to develop peptide-linked gold nanoparticles that have unique optical properties and can serve as beacons to visualize the apoptotic drug response in cancer cells at the single-molecule level. By thorough analyses of their trajectories, one can reveal early-stage caspase-3 activation in live cells continuously and with no ambiguity.

KEYWORDS:

Caspase; Gold nanoparticles; Live cells; Plasmon coupling; Single molecule

[Indexed for MEDLINE]

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