Skip to main content
SpringerLink
Log in

EPR Interfaced To Rapid Mixing

  • Chapter
Biomedical EPR, Part B: Methodology, Instrumentation, and Dynamics

Part of the book series: Biological Magnetic Resonance ((BIMR,volume 24/B))

Abstract

We describe the development of flow and stopped-flow EPR starting with large mixing devices designed for standard metallized EPR cavities and proceeding to micro mixing systems intimately attached to micro resonant structures. The characteristics of resonant structures dedicated to close attachment of liquid mixing systems with low dead volumes, short mixing times, and minimal microwave microphonic perturbation are outlined. The application of such systems to study of radical systems, either spin labelled, naturally occurring, or trapped, is reviewed, and recent experiments to probe rapid folding kinetics of site-directed spin labeled protein are summarized.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

7. References

  • Bonini, M. G., R. Radi, G. Ferrer-Sueta, A. M. Ferreira, and O. Augusto. 1999. Direct EPR detection of the carbonate radical anion produced from peroxynitrite and carbon dioxide. J Biol Chem 274:10802–10806.

    Article  PubMed  CAS  Google Scholar 

  • Bonini, M. G., and O. Augusto. 2001. Carbon Dioxide Stimulates The Production Of Thiyl, Sulfinyl, And Disulfide Radical Anion From Thiol Oxidation By Peroxynitrite. J Biol Chem 276:9749–9754.

    Article  PubMed  CAS  Google Scholar 

  • Borbat, P., and J. H. Freed. 2000. Double Quantum ESR and Distance Measurements. In: Biological Magnetic Resonance: Distance Measurements in Biological Systems by EPR. L. J. Berliner, G. R. Eaton, and S. S. Eaton, editors. Plenum, New York. 383–459.

    Google Scholar 

  • Borbat, P. P., H. S. Mchaourab, and J. H. Freed. 2002. Protein Structure Determination Using Long-Distance Constraints From Double-Quantum Coherence ESR: Study Of T4 Lysozyme. J Am Chem Soc 124:5304–5314.

    Article  PubMed  CAS  Google Scholar 

  • Borg, D. C. 1964a. Continuous Flow Methods Adapated For Epr Apparatus. In: Rapid Mixing and Sampling Techniques in Biochemistry. B. Chance, R. H. Eisenhardt, Q. H. Gibson, and K. K. Lonberg-Holm, editors. Academic Press, New York. 135–149.

    Google Scholar 

  • Borg, D. C. 1964b. An Improved Flow System For Electron Paramagnetic Resonance Spectrometry Of Aqueous Solutions. Nature 201:1087–1090.

    PubMed  CAS  Google Scholar 

  • Borg, D. C., and J. J. Elmore. 1967. Continuous Flow Apparatus For EPR Spectroscopy At 35 GHz. In: Magnetic Resonance in Biological Systems. A. Ehrenberg, B. G. Malmstrom, and T. Vanngard, editors. Pergamon Press, Oxford. 383–387.

    Google Scholar 

  • Brems, D. N., and E. Stellwagen. 1983. Manipulation Of The Observed Kinetic Phases In The Refolding Of Denatured Ferricytochromes C. J Biol Chem 258:3655–3660.

    PubMed  CAS  Google Scholar 

  • Bromberg, S. E., and I. Y. Chan. 1992. Enhanced Sensitivity For High-Pressure EPR Using Dielectric Resonators. Rev. Sci. Instrum. 63:3670–3676.

    Article  CAS  Google Scholar 

  • Buettner, G. R. 1987. Spin Trapping: ESR Parameters Of Spin Adducts. Free Rad. Biol. Med. 3:259–303.

    PubMed  CAS  Google Scholar 

  • Chance, B. 1940a. The Accelerated Flow Method For Rapid Reactions. Part I. Analysis. Journal of the Franklin Institute 229:455–476.

    CAS  Google Scholar 

  • Chance, B. 1940b. The Accelerated Flow Method For Rapid Reactions. Part II. Design, constructions, and tests. Journal of the Franklin Institute 229:737–766.

    CAS  Google Scholar 

  • Cohn, S. B. 1968. Microwave Bandpass Filters Containing High-Q Dielectric Resonators. IEEE Trans. Microwave Theory Tech 16:218–227.

    Google Scholar 

  • Colón, W., G. A. Elöve, L. P. Wakem, F. Sherman, and H. Roder. 1996. Side Chain Packing Of The N-And C-Terminal Helices Plays A Critical Role In The Kinetics Of Cytochrome C Folding. Biochemistry 35:5538–5549.

    PubMed  Google Scholar 

  • Colón, W., L. P. Wakem, F. Sherman, and H. Roder. 1997. Identification Of The Predominant Non-Native Histidine Ligand In Unfolded Cytochrome C. Biochemistry 36:12535–12541.

    PubMed  Google Scholar 

  • Creighton, T. E. 1994. The Energetic Ups And Downs Of Protein Folding [News]. Nat Struct Biol 1 :135–138.

    Article  PubMed  CAS  Google Scholar 

  • Davies, P. W. 1962. In: Physical Techniques in Biological Research. Academic Press, New York. 137–179.

    Google Scholar 

  • DeWeerd, K., V. Grigoryants, Y. Sun, J. S. Fetrow, and C. P. Scholes. 2001. EPR-Detected Folding Kinetics Of Externally Located Cysteine-Directed Spin-Labeled Mutants Of Iso-1-Cytochrome C. Biochemistry 40:15846–15855.

    Article  PubMed  CAS  Google Scholar 

  • Dykstra, R. W., and G. D. Markham. 1986. A Dielectric Sample Resonator For Enhanced Sensitivity Of EPR Spectroscopy. J. Mag. Reson. 69:350–355.

    CAS  Google Scholar 

  • Ebert, B., G. V. Semisotnov, and N. A.-Rodionova. 1990. Studies On Globular Protein Refolding Kinetics By ESR Stopped Flow Spectroscopy. Stud. Biophys. 137:125–132.

    CAS  Google Scholar 

  • Elöve, G. A., A. K. Bhuyan, and H. Roder. 1994. Kinetic Mechanism Of Cytochrome C Folding: Involvement Of The Heme And Its Ligands. Biochemistry 33:6925–6935.

    PubMed  Google Scholar 

  • Elöve, G. A., A. F. Chaffotte, H. Roder, and M. E. Goldberg. 1992. Early Steps In Cytochrome C Folding Probed By Time-Resolved Circular Dichroism And Fluorescence Spectroscopy. Biochemistry 31:6876–6883.

    PubMed  Google Scholar 

  • Englander, S. W., T. R. Sosnick, L. C. Mayne, M. Shtilerman, P. X. Qi, and Y. Bai. 1998. Fast And Slow Folding In Cytochrome C. Acc. Chem. Res. 31:737–744.

    Article  CAS  Google Scholar 

  • Fan, C., J. F. Bank, R. G. Dorr, and C. P. Scholes. 1988. An Electron Nuclear Double Resonance Investigation Of Redox-Induced Electronic Structural Change At Copper A 2+ In Cytochrome C Oxidase. J. Biol. Chem. 263:3588–3591.

    PubMed  CAS  Google Scholar 

  • Fenton, H. J. H. 1894. J. Chem. Soc. Trans. 65:899–910.

    Article  CAS  Google Scholar 

  • Fetrow, J. S., T. S. Cardillo, and F. Sherman. 1989. Deletions And Replacements Of Omega Loops In Yeast Iso-1-Cytochrome C. Proteins 6:372–381.

    Article  PubMed  CAS  Google Scholar 

  • Feynman, R. P., R. B. Leighton, and M. Sands. 1964. The Feynman Lectures on Physics. Addison-Wesley, Reading, MA.

    Google Scholar 

  • Froncisz, W., and J. S. Hyde. 1982. The Loop-Gap Resonator: A New Microwave Lumped Circuit ESR Sample Structure. J. Magn. Reson. 47:515–521.

    CAS  Google Scholar 

  • Froncisz, W., C. S. Lai, and J. S. Hyde. 1985. Spin-Label Oximetry: Kinetic Study Of Cell Respiration Using A Rapid-Passage T 1-Sensitive Electron Spin Resonance Display. Proc. Natl. Acad. Sci. U. S. A. 82:411–415.

    PubMed  CAS  Google Scholar 

  • Grigoryants, V. M., A. V. Veselov, and C. P. Scholes. 2000. Variable Velocity Liquid Flow EPR Applied To Submillisecond Protein Folding. Biophys J 78:2702–2708.

    PubMed  CAS  Google Scholar 

  • Hagen, S. J., J. Hofrichter, A. Szabo, and W. A. Eaton. 1996. Diffusion-Limited Contact Formation In Unfolded Cytochrome C: Estimating The Maximum Rate Of Protein Folding. Proc Natl Acad Sci USA 93:11615–11617.

    Article  PubMed  CAS  Google Scholar 

  • Halliwell, B., and J. M. C. Gutteridge. 1989. Free Radicals in Biology and Medicine. Second Edition. Clarendon Press, Oxford.

    Google Scholar 

  • Hubbell, W. L., and C. Altenbach. 1994. Investigation Of Structure And Dynamics In Membrane Proteins Using Site-Directed Spin Labeling. Curr. Opin. Struct. Biol. 4:566–573.

    Article  CAS  Google Scholar 

  • Hubbell, W. L., D. S. Cafiso, and C. Altenbach. 2000. Identifying Conformational Changes With Site-Directed Spin Labeling. Nat Struct Biol 7:735–739.

    Article  PubMed  CAS  Google Scholar 

  • Hubbell, W. L., W. Froncisz, and J. S. Hyde. 1987. Continuous And Stopped-Flow EPR Spectrometer Based On A Loop Gap Resonator. Rev. Sci. Instrum. 58:1879–1886.

    Article  Google Scholar 

  • Hubbell, W. L., A. Gross, R. Langen, and M. A. Lietzow. 1998. Recent Advances In Site-Directed Spin Labeling Of Proteins. Curr. Opin. Struct. Biol. 8:649–656.

    Article  PubMed  CAS  Google Scholar 

  • Jaworski, M., A. Sienkiewicz, and C. P. Scholes. 1997. Double-Stacked Dielectric Resonator For Sensitive EPR Measurements. J Magn Reson 124:87–96.

    Article  PubMed  CAS  Google Scholar 

  • Jiang, J. J., J. F. Bank, W. W. Zhao, and C. P. Scholes. 1992. The Method Of Time-Resolved Spin-Probe Oximetry. Its Application To Oxygen Consumption By Cytochrome c Oxidase. Biochemistry 31:1331–1339.

    Article  PubMed  CAS  Google Scholar 

  • Klimes, N., G. Lassmann, and B. Ebert. 1980. Time-Resolved EPR Spectroscopy. Stopped-Flow EPR Apparatus For Biological Application. J. Magn. Reson. 37:53–59.

    CAS  Google Scholar 

  • Lai, C. S., L. E. Hopwood, J. S. Hyde, and S. Lukiewicz. 1982. ESR Studies Of Oxygen Uptake By Chinese Hamster Ovary Cells During The Cell Cycle. Proc. Natl. Acad. Sci. U. S. A. 79:1166–1170.

    PubMed  CAS  Google Scholar 

  • Lassmann, G., L. Thelander, and A. Graslund. 1992. EPR Stopped-Flow Studies Of The Reaction Of The Tyrosyl Radical Of Protein R2 From Ribonucleotide Reductase With Hydroxyurea. Biochem Biophys Res Commun 188:879–887.

    Article  PubMed  CAS  Google Scholar 

  • Lassmann, G., L. A. Eriksson, F. Himo, F. Lendzian, and W. Lubitz. 1999. Electronic Structure Of A Transient Histidine Radical In Liquid Aqueous Solution: EPR Continuous-Flow Studies And Density Functional Calculations. J. Phys. Chem. A 103:1283–1290.

    Article  CAS  Google Scholar 

  • Lassmann, G., L. A. Eriksson, F. Lendzian, and W. Lubitz. 2000. Structure Of A Transient Neutral Histidine Radical In Solution: EPR Continuous-Flow Studies In A Ti 3+/EDTA-Fenton System And Density Functional Calculations. J. Phys. Chem. A 104:9144–9152.

    Article  CAS  Google Scholar 

  • Leszczynski, J. F., and G. D. Rose. 1986. Loops In Globular Proteins: A Novel Category Of Secondary Structure. Science 234:849–855.

    PubMed  CAS  Google Scholar 

  • Louie, G. V., and G. D. Brayer. 1990. High-Resolution Refinement Of Yeast Iso-1-Cytochrome c And Comparisons With Other Eukaryotic Cytochromes c. J Mol Biol 214:527–555.

    Article  PubMed  CAS  Google Scholar 

  • Marx, U., G. Lassmann, H. G. Holzhutter, D. Wustner, P. Muller, A. Hohlig, J. Kubelt, and A. Herrmann. 2000. Rapid Flip-Flop Of Phospholipids In Endoplasmic Reticulum Membranes Studied By A Stopped-Flow Approach. Biophys J:2628–2640.

    Google Scholar 

  • Marx, U., G. Lassmann, K. Wimalasena, P. Muller, and A. Herrmann. 1997. Rapid Kinetics Of Insertion And Accessibility Of Spin-Labeled Phospholipid Analogs In Lipid Membranes: A Stopped-Flow Electron Paramagnetic Resonance Approach. Biophys J 73:1645–1654.

    Article  PubMed  CAS  Google Scholar 

  • Mascarenhas, R., Y. H. Wei, C. P. Scholes, and T. E. King. 1983. Interaction In Cytochrome c Oxidase Between Cytochrome a3 Ligated With Nitric Oxide And Cytochrome a. J. Biol. Chem. 258:5348–5351.

    PubMed  CAS  Google Scholar 

  • Morar, A. S., D. Kakouras, G. B. Young, J. Boyd, and G. J. Pielak. 1999. Expression Of 15 N-Labeled Eukaryotic Cytochrome c In Escherichia Coli. J Biol Inorg Chem 4:220–222.

    PubMed  CAS  Google Scholar 

  • Nall, B. T., and T. A. Landers. 1981. Guanidine Hydrochloride Induced Unfolding Of Yeast Iso-2 Cytochrome c. Biochemistry 20:5403–5411.

    PubMed  CAS  Google Scholar 

  • Osterhout, J. J., Jr., and B. T. Nall. 1985. Slow refolding kinetics in yeast iso-2 cytochrome c. Biochemistry 24:7999–8005.

    PubMed  CAS  Google Scholar 

  • Plurde, K. J., D, F. Linn, H. M. O’Bryan, Jr., and J. Thomson, Jr. 1975. Ba 2 Ti 9 O 20 as a Microwave Dielectric Resonator. J. Am. Ceram. Soc. 58:418–427.

    Google Scholar 

  • Plurde, K. J., and C.-L. Ren. 1981. Application Of Dielectric Resonators In Microwave Components. IEEE Trans. Microwave Theory Tech 29:754–770.

    Google Scholar 

  • Pollock, W. B., F. I. Rosell, M. B. Twitchett, M. E. Dumont, and A. G. Mauk. 1998. Bacterial expression of a mitochondria] cytochrome c. Trimethylation Of Lys72 In Yeast Iso-1-Cytochrome C And The Alkaline Conformational Transition. Biochemistry 37:6124–6131.

    Article  PubMed  CAS  Google Scholar 

  • Qu, K., J. L. Vaughn, A. Sienkiewicz, C. P. Scholes, and J. S. Fetrow. 1997. Kinetics And Motional Dynamics Of Spin-Labeled Yeast Iso-1-Cytochrome c: 1. Stopped-Flow Electron Paramagnetic Resonance As A Probe For Protein Folding/Unfolding Of The CTerminal Helix Spin-Labeled At Cysteine 102. Biochemistry 36:2884–2897.

    Article  PubMed  CAS  Google Scholar 

  • Regenfuss, P., R. M. Clegg, M. J. Fulwyler, F. J. Barrentes, and T. M. Jovin. 1985. Mixing Liquids In Microseconds. Rev. Sci. Instrum. 56:283–290.

    Article  CAS  Google Scholar 

  • Richtmyer, D. 1939. Dielectric Resonators. J. Appl. Physics 10:391–398.

    Article  Google Scholar 

  • Roder, H., G. A. Elöve, and S. W. Englander. 1988. Structural Characterization Of Folding Intermediates In Cytochrome C By H-Exchange Labelling And Proton NMR. Nature 335:700–704.

    Article  PubMed  CAS  Google Scholar 

  • Sahlin, M., G. Lassmann, S. Potsch, B. M. Sjoberg, and A. Graslund. 1995. Transient Free Radicals In Iron/Oxygen Reconstitution Of Mutant Protein R2 Y122F. Possible Participants In Electron Transfer Chains In Ribonucleotide Reductase. J Biol Chem 270:12361–12372.

    PubMed  CAS  Google Scholar 

  • Shastry, M. C., and H. Roder. 1998. Evidence For Barrier-Limited Protein Folding Kinetics On The Microsecond Time Scale. Nat Struct Biol 5:385–392.

    Article  PubMed  CAS  Google Scholar 

  • Shastry, M. C. R., J. M. Sauder, and H. Roder. 1998. Kinetic And Structural Analysis Of Submillisecond Folding Events In Cytochrome c. Acc. Chem. Res. 31:717–725.

    CAS  Google Scholar 

  • Sienkiewicz, A., A. M. da Costa Ferreira, B. Danner, and C. P. Scholes. 1999. Dielectric Resonator-Based Flow And Stopped-Flow EPR With Rapid Field Scanning: A Methodology For Increasing Kinetic Information. J Magn Reson 136:137–142.

    Article  PubMed  CAS  Google Scholar 

  • Sienkiewicz, A., K. Qu, and C. P. Scholes. 1994. Dielectric Resonator-Based Stopped-Flow EPR. Rev. Sci. Instrum. 65:68–74.

    Article  CAS  Google Scholar 

  • Sosnick, T. R., L. Mayne, R. Hiller, and S. W. Englander. 1994. The Barriers In Protein Folding. Nat Struct Biol 1:149–156.

    Article  PubMed  CAS  Google Scholar 

  • Sosnick, T. R., M. D. Shtilerman, L. Mayne, and S. W. Englander. 1997. Ultrafast Signals In Protein Folding And The Polypeptide Contracted State. Proc Natl Acad Sci U S A 94:8545–8550.

    Article  PubMed  CAS  Google Scholar 

  • Stevens, T. H., C. T. Martin, H. Wang, G. W. Brudvig, C. P. Scholes, and S. I. Chan. 1982. The Nature Of CuA In Cytochrome c Oxidase. J Biol Chem 257:12106–12113.

    PubMed  CAS  Google Scholar 

  • Tsong, T. Y. 1976. Ferricytochrome c Chain Folding Measured By The Energy Transfer Of Tryptophan 59 To The Heme Group. Biochemistry 15:5467–5473.

    Article  PubMed  CAS  Google Scholar 

  • Tullius, T. D. 1988. DNA Footprinting With Hydroxyl Radical. Nature 332:663–664.

    Article  PubMed  CAS  Google Scholar 

  • van Camp, H. L., Y. H. Wei, C. P. Scholes, and T. E. King. 1978. Electron Nuclear Double Resonance Of Cytochrome Oxidase: Nitrogen And Proton ENDOR From The ‘Copper’ EPR Signal. Biochim Biophys Acta 537:238–246.

    PubMed  Google Scholar 

  • Vanderkooi, J. M., G. Maniara, T. J. Green, and D. F. Wilson. 1987. An Optical Method For Measurement Of Dioxygen Concentration Based Upon Quenching Of Phosphorescence. J Biol Chem 262:5476–5482.

    PubMed  CAS  Google Scholar 

  • Walsh, W. M., and L. W. Rupp. 1986. Enhanced ESR Sensitivity Using A Dielectric Resonator. Rev. Sci. Instrum. 57:2278–2279.

    Article  CAS  Google Scholar 

  • Wilson, D. F., W. L. Rumsey, T. J. Green, and J. M. Vanderkooi. 1988. The Oxygen Dependence Of Mitochondrial Oxidative Phosphorylation Measured By A New Optical Method For Measuring Oxygen Concentration. J Biol Chem 263:2712–2718.

    PubMed  CAS  Google Scholar 

  • Windrem, D. A., And W. Z. Plachy. 1980. The diffusion-solubility of oxygen in lipid bilayers. Biochim Biophys Acta 600:655–665.

    PubMed  CAS  Google Scholar 

  • Winkler, J. R., and H. B. Gray. 1998. Protein Folding. Acc. Chem. Res. 31:698.

    Article  CAS  Google Scholar 

  • Yamazaki, I., H. S. Mason, and L. Piette. 1960. Identification by Electron Paramagnetic Resonance Spectroscopy, Of Free Radicals Generated From Substrates By Peroxidase. J. Biol. Chem. 235:2444–2449.

    PubMed  CAS  Google Scholar 

  • Yamazaki, I., and L. H. Piette. 1990. ESR Spin-Trapping Studies On The Reaction Of Iron(2+) Ions With Hydrogen Peroxide-Reactive Species In Oxygen Toxicity In Biology. J. Biol. Chem. 265:13589–13594.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Chemistry, University at Albany — State University of New York, Albany, NY, 12222

    Charles P. Scholes

Authors
  1. Charles P. Scholes
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Denver, Denver, Colorado

    Sandra R. Eaton , Gareth R. Eaton  & Lawrence J. Berliner ,  & 

Rights and permissions

Reprints and permissions

Copyright information

© 2005 Kluwer Academic/Plenum Publishers

About this chapter

Cite this chapter

Scholes, C.P. (2005). EPR Interfaced To Rapid Mixing. In: Eaton, S.R., Eaton, G.R., Berliner, L.J. (eds) Biomedical EPR, Part B: Methodology, Instrumentation, and Dynamics. Biological Magnetic Resonance, vol 24/B. Springer, Boston, MA. https://doi.org/10.1007/0-306-48533-8_3

Download citation

  • DOI: https://doi.org/10.1007/0-306-48533-8_3

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-0-306-48532-9

  • Online ISBN: 978-0-306-48533-6

  • eBook Packages: MedicineMedicine (R0)

Publish with us

Policies and ethics

Search

Navigation