A photoenhanced oxidation of amino acids and the cross-linking of lysozyme mediated by tetrazolium salts.

Tetrazolium salts (TZs) are pervasively utilized as precursors in the dye industry, colorimetric probes in enzyme assays and for exploring nanomaterial toxicity, but its own toxicity is not investigated enough so far. Using femtosecond transient absorption spectroscopy, nanosecond pulse radiolysis (ns-PRL), western blotting and UV-vis absorption spectroscopy, here we characterized a neutral tetrazolinyl radical (with the same maximum absorption at 420 nm and different lifetimes of 5.0 and 9.0 μs for two selected TZs), the key intermediate of TZs reduction, and noticed TZs-formazan production under UV light irradiation accompanied by 41% increase in the cross-linking of lysozyme (Lyso, model protein) compared to TZs-free sample, which uncovered the photoenhanced oxidation of TZs towards Lyso. The ns-PRL in a reductive atmosphere simulated the electron/proton donors of amino acid residues in Lyso upon photoexcitation and revealed the reduction mechanism of TZs, as that first followed one-electron-transfer and then probably proton-coupled electron transfer. This is the first time to report on the photoenhanced oxidation mechanism of TZs, which would provide new insights into the applications of TZs in cell biology, "click" chemistry and nanotoxicology.

[1]  P. Hägglund,et al.  Photo-oxidation of lysozyme triggered by riboflavin is O2-dependent, occurs via mixed type 1 and type 2 pathways, and results in inactivation, site-specific damage and intra- and inter-molecular crosslinks. , 2020, Free radical biology & medicine.

[2]  P. Di Mascio,et al.  Singlet oxygen induced protein aggregation: lysozyme cross-link formation and nLC-MS/MS characterization. , 2019, Journal of mass spectrometry : JMS.

[3]  E. Ermakova,et al.  Alkyl triphenylphosphonium surfactants as nucleic acid carriers: complexation efficacy toward DNA decamers, interaction with lipid bilayers and cytotoxicity studies. , 2019, Physical chemistry chemical physics : PCCP.

[4]  Mariana P Serrano,et al.  Evidence of the effectiveness of Resveratrol in the prevention of guanine one-electron oxidation: possible benefits in cancer prevention. , 2019, Physical chemistry chemical physics : PCCP.

[5]  C. Menck,et al.  The key role of UVA‐light induced oxidative stress in human Xeroderma Pigmentosum Variant cells , 2019, Free radical biology & medicine.

[6]  P. Sandhu,et al.  Attenuation of neuroblastoma cell growth by nisin is mediated by modulation of phase behavior and enhanced cell membrane fluidity. , 2019, Physical chemistry chemical physics : PCCP.

[7]  R. Swaminathan,et al.  Protein charge transfer absorption spectra: an intrinsic probe to monitor structural and oligomeric transitions in proteins. , 2017, Faraday discussions.

[8]  Jingye（李景烨） Li,et al.  Photo-enhanced oxidizability of tetrazolium salts and its impact on superoxide assaying. , 2016, Chemical communications.

[9]  P. Di Mascio,et al.  Production of lysozyme and lysozyme-superoxide dismutase dimers bound by a ditryptophan cross-link in carbonate radical-treated lysozyme. , 2015, Free radical biology & medicine.

[10]  Jingye（李景烨） Li,et al.  Graphene Oxide Transparent Hybrid Film and Its Ultraviolet Shielding Property. , 2015, ACS applied materials & interfaces.

[11]  R. Longo,et al.  Synthesis and structure of 1-benzyl-5-amino-1H-tetrazole in the solid state and in solution: Combining X-ray diffraction, 1H NMR, FT–IR, and UV–Vis spectra and DFT calculations , 2015 .

[12]  Y. Kitagawa,et al.  Accurate standard hydrogen electrode potential and applications to the redox potentials of vitamin C and NAD/NADH. , 2015, The journal of physical chemistry. A.

[13]  R. Hurt,et al.  Antioxidant chemistry of graphene-based materials and its role in oxidation protection technology. , 2014, Nanoscale.

[14]  Pedro J J Alvarez,et al.  Photochemical transformation of carboxylated multiwalled carbon nanotubes: role of reactive oxygen species. , 2013, Environmental science & technology.

[15]  Jing Kong,et al.  Antibacterial activity of graphite, graphite oxide, graphene oxide, and reduced graphene oxide: membrane and oxidative stress. , 2011, ACS nano.

[16]  K. Akiyoshi,et al.  Construction of protein-crosslinked nanogels with vitamin B6 bearing polysaccharide , 2011 .

[17]  H. Gray,et al.  Tryptophan-Accelerated Electron Flow Through Proteins , 2008, Science.

[18]  W. Dong,et al.  Investigation of the reactivity of hydrated electron toward perfluorinated carboxylates by laser flash photolysis , 2007 .

[19]  T. Ohsaka,et al.  Electrochemical and spectroscopic studies on electron-transfer reaction between novel water-soluble tetrazolium salts and a superoxide ion , 2004 .

[20]  P. Jollès,et al.  Re-evaluation of intramolecular long-range electron transfer between tyrosine and tryptophan in lysozymes. Evidence for the participation of other residues. , 2003, European journal of biochemistry.

[21]  Peter Eck,et al.  Vitamin C as an Antioxidant: Evaluation of Its Role in Disease Prevention , 2003, Journal of the American College of Nutrition.

[22]  K. Sharpless,et al.  A click chemistry approach to tetrazoles by Huisgen 1,3-dipolar cycloaddition: synthesis of 5-sulfonyl tetrazoles from azides and sulfonyl cyanides. , 2002, Angewandte Chemie.

[23]  S. Yao,et al.  Kinetic observation of rapid electron transfer between pyrimidine electron adducts and sensitizers of riboflavin, flavin adenine dinucleotide (FAD) and chloranil: a pulse radiolysis study , 2000 .

[24]  W. Mclaughlin,et al.  Radiolytic reactions of nitro blue tetrazolium under oxidative and reductive conditions: a pulse radiolysis study , 1999 .

[25]  H. Ukeda,et al.  Flow-Injection Assay of Superoxide Dismutase Based on the Reduction of Highly Water-Soluble Tetrazolium , 1999 .

[26]  H. Ukeda,et al.  Spectrophotometric assay for superoxide dismutase based on tetrazolium salt 3'--1--(phenylamino)-carbonyl--3, 4-tetrazolium]-bis(4-methoxy-6-nitro)benzenesulfonic acid hydrate reduction by xanthine-xanthine oxidase. , 1997, Analytical biochemistry.

[27]  M. Sutherland,et al.  The tetrazolium dyes MTS and XTT provide new quantitative assays for superoxide and superoxide dismutase. , 1997, Free radical research.

[28]  K. Bobrowski,et al.  Pulse radiolysis studies of intramolecular electron transfer in model peptides and proteins. 7. Trp-->TyrO radical transformation in hen egg-white lysozyme. Effects of pH, temperature, Trp62 oxidation and inhibitor binding. , 1997, Biophysical chemistry.

[29]  M. Ishiyama,et al.  A combined assay of cell viability and in vitro cytotoxicity with a highly water-soluble tetrazolium salt, neutral red and crystal violet. , 1996, Biological & pharmaceutical bulletin.

[30]  M. Čeppan,et al.  Radical intermediates in the redox reactions of tetrazolium salts in aprotic solvents (cyclovoltammetric, EPR and UV-VIS study). , 1994, Free radical research.

[31]  D. Njus,et al.  Vitamins C and E donate single hydrogen atoms in vivo , 1991, FEBS letters.

[32]  Z. Alfassi,et al.  Long range electron transfer between tyrosine and tryptophan in hen egg-white lysozyme. , 1991, Biochimica et biophysica acta.

[33]  M. Klapper,et al.  Pulse radiolytic measurement of redox potentials: the tyrosine and tryptophan radicals. , 1989, Biochemistry.

[34]  C. G. Atkins,et al.  Two-photon dissociation of H2O at 266 nm , 1988 .

[35]  B. Ames,et al.  Antioxidant activity of carnosine, homocarnosine, and anserine present in muscle and brain. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[36]  A. Ross,et al.  Reactivity of HO2/O−2 Radicals in Aqueous Solution , 1985 .

[37]  L. K. Patterson,et al.  Direct observation of monophotonic photoionization in tryptophan excited by 300-nm radiation. A laser photolysis study , 1983 .

[38]  R. J. Robbins,et al.  Photophysics of Aqueous Tryptophan: pH and Temperature Effects , 1980 .

[39]  S. Venitt,et al.  Bacterial mutagenicity tests of phenazine methosulphate and three tetrazolium salts. , 1979, Mutation research.

[40]  M. Karel,et al.  Changes in lysozyme due to reactions with peroxidizing methyl linoleate in a dehydrated model system. , 1976, Journal of agricultural and food chemistry.

[41]  R. G. Butcher,et al.  Studies on the reduction of tetrazolium salts , 1973, Histochemie.

[42]  F. Richards,et al.  Glutaraldehyde as a protein cross-linking reagent , 1968 .

[43]  M. Singla,et al.  Coupling of chromatographic analyses with pretreatment for the determination of bioactive compounds in Emblica officinalis juice , 2014 .

[44]  H. Freeman,et al.  Synthetic dyes based on environmental considerations. Part 2: Iron complexes formazan dyes☆ , 1996 .

[45]  E. Silva,et al.  Isolation and photo-oxidation of lysozyme fragments , 1981, Radiation and environmental biophysics.