Detection of single-molecule H2O2 signalling from epidermal growth factor receptor using fluorescent single-walled carbon nanotubes.
An emerging concept in cell signalling is the natural role of reactive oxygen species such as hydrogen peroxide (H2O2) as beneficial messengers in redox signalling pathways. The nature of H2O2 signalling is confounded, however, by difficulties in tracking it in living systems, both spatially and temporally, at low concentrations. Here, we develop an array of fluorescent single-walled carbon nanotubes that can selectively record, in real time, the discrete, stochastic quenching events that occur as H2O2 molecules are emitted from individual human epidermal carcinoma cells stimulated by epidermal growth factor. We show mathematically that such arrays can distinguish between molecules originating locally on the cell membrane from other contributions. We find that epidermal growth factor induces 2 nmol H2O2 locally over a period of 50 min. This platform promises a new approach to understanding the signalling of reactive oxygen species at the cellular level.
Jin
Hong
H
Department of Chemical Engineering, Massachusetts Institute of Technology, Building 66-566, 77 Massachusetts Ave, Cambridge, Massachusetts 02139-4307, USA.
Heller
Daniel A
DA
Kalbacova
Marie
M
Kim
Jong-Ho
JH
Zhang
Jingqing
J
Boghossian
Ardemis A
AA
Maheshri
Narendra
N
Strano
Michael S
MS
D015374:Biosensing Techniques / Q000379:methods*; D002450:Cell Communication; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry / Q000378:metabolism*; D008390:Markov Chains; D009010:Monte Carlo Method; D037742:Nanotubes, Carbon / Q000737:chemistry*; D015398:Signal Transduction; D013269:Stochastic Processes
D005456:Fluorescent Dyes; D037742:Nanotubes, Carbon; D006861:Hydrogen Peroxide; D066246:ErbB Receptors