Detection of single-molecule H2O2 signaling 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.

[1]  T. J. Hardwick.  THE RATE CONSTANT OF THE REACTION BETWEEN FERROUS IONS AND HYDROGEN PEROXIDE IN ACID SOLUTION , 1957 .

[2]  W. Pryor.  Free Radicals in Biology , 1976 .

[3]  C. Foote.  CHAPTER 3 – Photosensitized Oxidation and Singlet Oxygen: Consequences in Biological Systems , 1976 .

[4]  I. Fridovich.  The biology of oxygen radicals. , 1978, Science.

[5]  S. L. Issler,et al.  Involvement of the azide radical in the quenching of singlet oxygen by azide anion in water , 1982 .

[6]  S. Cohen,et al.  Epidermal growth factor , 1972, The Journal of investigative dermatology.

[7]  S. Linn,et al.  Toxic DNA damage by hydrogen peroxide through the Fenton reaction in vivo and in vitro. , 1988, Science.

[8]  A. Ullrich,et al.  Functional analysis of the ligand binding site of EGF‐receptor utilizing chimeric chicken/human receptor molecules. , 1989, The EMBO journal.

[9]  M. Baerns,et al.  Oxidative coupling of methane in the gas phase. Kinetic simulation and experimental verification , 1990 .

[10]  Barry Halliwell,et al.  DNA damage by oxygen‐derived species Its mechanism and measurement in mammalian systems , 1991, FEBS letters.

[11]  J. Mendelsohn,et al.  Consumption of EGF by A431 cells: evidence for receptor recycling , 1993, The Journal of cell biology.

[12]  Application of sensitivity analysis to the reduction of a complex kinetic model for the homogeneous oxidative coupling of methane , 1993 .

[13]  M. Kasha,et al.  Singlet molecular oxygen in the Haber-Weiss reaction. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[14]  E. Okabe,et al.  Inhibition by singlet molecular oxygen of the vascular reactivity in rabbit mesenteric artery , 1997, British journal of pharmacology.

[15]  R. Haugland,et al.  A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. , 1997, Analytical biochemistry.

[16]  E. Tekle,et al.  Epidermal Growth Factor (EGF)-induced Generation of Hydrogen Peroxide , 1997, The Journal of Biological Chemistry.

[17]  Jia-Ling Yang,et al.  Singlet oxygen is the major species participating in the induction of DNA strand breakage and 8‐hydroxydeoxyguanosine adduct by lead acetate , 1999, Environmental and molecular mutagenesis.

[18]  R. Lerner,et al.  Antibodies have the intrinsic capacity to destroy antigens. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Y. Yarden,et al.  Untangling the ErbB signalling network , 2001, Nature Reviews Molecular Cell Biology.

[20]  R. Lerner,et al.  Antibody Catalysis of the Oxidation of Water , 2001, Science.

[21]  J. Imlay,et al.  Pathways of oxidative damage. , 2003, Annual review of microbiology.

[22]  H. Kovacic,et al.  Monolayer versus aggregate balance in survival process for EGF‐induced apoptosis in A431 carcinoma cells: Implication of ROS‐P38 MAPK‐integrin α2β1 pathway , 2004, International journal of cancer.

[23]  Jorge Nieva,et al.  The antibody-catalyzed water oxidation pathway--a new chemical arm to immune defense? , 2004, Trends in biochemical sciences.

[24]  Roy S Herbst,et al.  Review of epidermal growth factor receptor biology. , 2004, International journal of radiation oncology, biology, physics.

[25]  Sung Ho Ryu,et al.  Sequential Activation of Phosphatidylinositol 3-Kinase, βPix, Rac1, and Nox1 in Growth Factor-Induced Production of H2O2 , 2004, Molecular and Cellular Biology.

[26]  Demianova Yulia.  Hydrogen peroxide generated extracellularly by receptor-ligand interaction facilitates cell signaling , 2005 .

[27]  G. K. Budnikov,et al.  Reactions of superoxide anion radical with antioxidants and their use in voltammetry , 2005 .

[28]  D. Golde,et al.  Hydrogen peroxide generated extracellularly by receptor-ligand interaction facilitates cell signaling. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[29]  S. Lukyanov,et al.  Genetically encoded fluorescent indicator for intracellular hydrogen peroxide , 2006, Nature Methods.

[30]  W. R. Taylor,et al.  In vivo imaging of hydrogen peroxide with chemiluminescent nanoparticles. , 2007, Nature materials.

[31]  D. Velegol,et al.  Chemotaxis of nonbiological colloidal rods. , 2007, Physical review letters.

[32]  J. Tour,et al.  Stepwise Quenching of Exciton Fluorescence in Carbon Nanotubes by Single-Molecule Reactions , 2007, Science.

[33]  J. Lambert,et al.  Measuring the lifetime of singlet oxygen in a single cell: addressing the issue of cell viability , 2007, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[34]  Evan W. Miller,et al.  Molecular imaging of hydrogen peroxide produced for cell signaling. , 2007, Nature chemical biology.

[35]  Nicholas A. Ramey,et al.  Imaging Mitochondria in Living Corneal Endothelial Cells Using Autofluorescence Microscopy , 2007, Photochemistry and photobiology.

[36]  B. Morgan,et al.  Hydrogen Peroxide Sensing and Signaling , 2022 .

[37]  Zhuang Liu,et al.  Selective probing and imaging of cells with single walled carbon nanotubes as near-infrared fluorescent molecules. , 2008, Nano letters.

[38]  D. Shaw,et al.  Superoxide dismutase 1 (SOD1) is essential for H2O2-mediated oxidation and inactivation of phosphatases in growth factor signaling , 2008, Proceedings of the National Academy of Sciences.

[39]  J. Imlay.  Cellular defenses against superoxide and hydrogen peroxide. , 2008, Annual review of biochemistry.

[40]  Michael S Strano,et al.  Stochastic analysis of stepwise fluorescence quenching reactions on single-walled carbon nanotubes: single molecule sensors. , 2008, Nano letters.

[41]  M. Strano,et al.  The rational design of nitric oxide selectivity in single-walled carbon nanotube near-infrared fluorescence sensors for biological detection. , 2009, Nature chemistry.

[42]  M. Kuimova,et al.  Singlet oxygen in a cell: spatially dependent lifetimes and quenching rate constants. , 2009, Journal of the American Chemical Society.

[43]  Michael S Strano,et al.  Multimodal optical sensing and analyte specificity using single-walled carbon nanotubes. , 2009, Nature nanotechnology.

[44]  J. Boilot,et al.  Single europium-doped nanoparticles measure temporal pattern of reactive oxygen species production inside cells. , 2009, Nature nanotechnology.