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Two dimensional diffusion of small molecules on protein surfaces: an EPR study of the restricted translational diffusion of protein-bound spin labels

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Abstract

Heisenberg spin exchange rates and dipole-dipole spin lattice relaxation rates for deuterated 14N- and 15N-spin labels bound selectively to the histidine His15 and to the lysines Lys13, 96, 97 of the lysozyme molecule have been determined with the aid of electron spin resonance spectroscopy. The results can be interpreted in terms of a two dimensional translational diffusion of the nitroxide tips of the spin labels along the protein surface within restricted surface areas. The spin labels are regarded as models for long amino acid side chains and as probes for the dynamics of protein and water in the vicinity of the protein surface. The translational diffusion coefficient DPII is reduced by a factor of between six and thirty compared to the value of D found for the spin labels in bulk water, its value for T = 295 K is given by (1.3±0.6)·10−10m2s−1 ≥ DΠ ≥ (2.4±0.3) 10−11 m2s−1.

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References

  • Abragam A (1970) The principles of nuclear magnetism. Clarendon Press, London Glasgow New York

    Google Scholar 

  • Brändle R, Krüger GJ, Müller-Warmuth W (1970) Impulsspektroskopische Untersuchungen der Elektronenspinrelaxation in freien Radikalen. Z Naturforsch 25A:1–11

    Google Scholar 

  • Case DA, Karplus M (1979) Dynamics of ligand binding to heme proteins. J Mol Biol 132:343–368

    Google Scholar 

  • Chandrasekhar S (1943) Stochastic problems in physics and astronomy. Rev Mod Phys 15:1–89

    Google Scholar 

  • Chothia C (1974) Hydrophobic bonding and accessible surface area in proteins. Nature 248:338–339

    Google Scholar 

  • Ciecierska-Tworek Z, Pong Van S, Griffith H (1973) Electron-electron dipolar splitting anisotropy of a dinitroxide oriented in a cristalline matrix. J Mol Struct 16:139–148

    Google Scholar 

  • Dombrowsky O (1990) Messungen der longitudinalen Elektronen-relaxationszeit an spinmarkierten Proteinen und niedermolekularen Modellsystemen. Dissertation, Ruhr-Universität, Bochum

    Google Scholar 

  • Dubin SB, Clark NA, Benedek GB (1971) Measurement of the rotational diffusion coefficient of lysozyme by depolarized light scattering: configuration of lysozyme in solution. J Chem Phys 54:5158–5164

    Google Scholar 

  • Eastman MP, Kooser RG, Das MR, Freed JH (1969) Studies of Heisenberg spin exchange in ESR spectra. I. Linewidth and saturation effects. J Chem Phys 51:2690–2709

    Google Scholar 

  • Eastman MP, Bruno GV, Freed JH (1970) ESR studies of Heisenberg spin exchange. II. Effects of radical charge and size. J Chem Phys 52:2511–2522

    Google Scholar 

  • Feix JP, Popp CA, Venkataramu SD, Beth AH, Park JH, Hyde JS (1984) An electron-electron double-resonance study of interactions between 14N- and 15N-stearic acid spin-label pairs: Lateral diffusion and vertical fluctuations in dimyristoylphosphatidylcholine. Biochemistry 23:2293–2299

    Google Scholar 

  • Frauenfelder H, Gratton E (1986) Protein dynamics and hydration. Methods Enzymol 127:207–216

    Google Scholar 

  • Freed JH (1976) Theory of slow tumbling ESR spectra for nitroxides. In: Berliner LJ (ed) Spin labeling theory and applications. Academic Press, New York, pp 53–132

    Google Scholar 

  • Gaffney BJ (1976) The chemistry of spin labels. In: Berliner LJ (ed) Spin labeling. Theory and applications. Academic Press, New York, pp 183–232

    Google Scholar 

  • Huisjen M, Hyde J (1974) A pulsed EPR spectrometer. Rev Sci Instrum 45:669–675

    Google Scholar 

  • Hwang LP, Freed JH (1975) Dynamic effects of pair correlation functions on spin relaxation by translational diffusion in liquids. J Chem Phys 63:4017–4025

    Google Scholar 

  • Hyde JS, Feix JB (1989) Electron-electron double resonance. In: Berliner LJ, Reuben J (ed) Spin labeling — theory and applications. Plenum Press, New York, pp 305–337

    Google Scholar 

  • Jolicoeur C, Friedman HL (1971) Effects of hydrophobic interactions on dynamics in aqueous solutions studied by EPR. Ber Bunsen-Gesell 75:248–257

    Google Scholar 

  • Kivelson D (1972) Electron relaxation in liquids. Selected topics. In: Muus LT, Atkins PW (eds) Electron spin relaxation in liquids. Plenum Press, New York London, pp 213–277

    Google Scholar 

  • Kooser RG, Volland WV, Freed JH (1969) ESR relaxation studies on orbitally degenerate free radicals. I. Benzene anion and tropenyl. J Chem Phys 50:5243–5257

    Google Scholar 

  • Korb JP, Torney DC, McConnell HM (1983) Dipolar correlation function and motional narrowing in finite two-dimensional spin systems. J Chem Phys 78:5782–5789

    Google Scholar 

  • Korb JP, Winterhalter M, McConnell HM (1984) Theory of spin relaxation by translational diffusion in two-dimensional systems. J Chem Phys 80:1059–1068

    Google Scholar 

  • Korb JP, Ahadi M, Zientara GP, Freed JH (1987) Dynamic effects of pair correlation functions of spin relaxation by translational diffusion in two-dimensional fluids. J Chem Phys 86:1125–1130

    Google Scholar 

  • Krüger GJ (1969) Magnetische Relaxation durch Translationsdiffusion in Flüssigkeiten. Z Naturforsch 21a:560–565

    Google Scholar 

  • Krüger GJ (1972) Electron spin relaxation in solutions of organic free radicals. Adv Mol Relax Process 3:235–251

    Google Scholar 

  • Lee S, Karplus M (1987) Kinetics of diffusion-influenced bimolecular reactions in solution. II. Effects of the gating mode and orientation-dependent reactivity. J Chem Phys 86:1904–1921

    Google Scholar 

  • Likhtenshtein GI (1976) Spin labeling methods in molecular biology. Wiley, New York London Sydney

    Google Scholar 

  • Likhtenshtein GI, Akhmedov YD, Ivanov LV Krinitskaya LA, Kokhanov YV (1974) Investigation of the lysozyme macromolecule by a spin-labeling method. Mol Biol 8:40–48

    Google Scholar 

  • Nayeem A, Rananavare SB, Sastry VSS, Freed JH (1989) Heisenberg spin exchange and molecular diffusion in liquid crystals. J Chem Phys 91:6887–6905

    Google Scholar 

  • Percival PW Hyde JS (1976) Saturation-recovery measurements of the spin-lattice relaxation times of some nitroxides in solution. J Magn Res 23:249–257

    Google Scholar 

  • Perutz MF, Mathews FS (1966) An X-ray study of azide methaemoglobin. J Mol Biol 21:199–202

    Google Scholar 

  • Polnaszek CF, Bryant RG (1984) Nitroxide radical induced solvent proton relaxation: Measurement of localized translational diffusion. J Chem Phys 81:4038–4045

    Google Scholar 

  • Popp CA, Hyde JS (1981) Effects of oxygen on ESR spectra of nitroxide spin-label probes of model membranes. J Magn Res 43:249–258

    Google Scholar 

  • Prabhananda BS, Hyde JS (1986) Study of molecular motions in liquids by electron spin relaxation: Halogenated p-semiquinone anions in alcohols. J Chem Phys 85:6705–6712

    Google Scholar 

  • Redhardt A, Daseler W (1987) A new method for absolute determination of radical concentration by EPR. J Biochem Biophys Methods 15:71–84

    Google Scholar 

  • Robinson B, Thomann H, Beth A, Fajer P, Dalton L (1985) The phenomenon of magnetic resonance: Theoretical considerations. In: Dalton LR (ed) EPR and advanced EPR studies of biological systems. CRC Press, Boca Raton, Florida, pp 11–111

    Google Scholar 

  • Rozantsev EG (1970) Free nitroxyl radicals. Plenum Press, New York London

    Google Scholar 

  • Sachse JH, King MD, Marsh D (1987) ESR determination of lipid translational diffusion coefficients at low spin-label concentrations in biological membranes, using exchange broadening, exchange narrowing, and dipole-dipole interactions. J Magn Res 71:385–404

    Google Scholar 

  • Schmidt PG, Kuntz ID (1984) Distance measurements in spin-labeled lysozyme. Biochemistry 23:4261–4266

    Google Scholar 

  • Schneider DJ, Freed JH (1989) Calculating slow motional magnetic resonance spectra. In: Berliner LJ, Reuben J (ed) Spin labeling —theory and applications. Plenum Press, New York, pp 1–76

    Google Scholar 

  • Steinhoff HJ (1988) A simple method for determination of rotational correlation times and separation of rotational and polarity effects from EPR spectra of spin-labeled biomolecules in a wide correlation time range. J Biochem Biophys Methods 17:237–248

    Google Scholar 

  • Steinhoff HJ (1990) Residual motion of hemoglobin-bound spin labels and protein dynamics: viscosity dependence of the rotational correlation times. Eur Biophys J 18:7–62

    Google Scholar 

  • Steinhoff HJ, Lieutenant K, Schlitter J (1989) Residual motion of hemoglobin-bound spin labels as a probe for protein dynamics. Z Naturforsch 44c:280–288

    Google Scholar 

  • Tabony J, Korb JP (1985) Interpretation of proton N.M.R. spin lattice relaxation time minima in heterogeneous systems by the effects of bounded two dimensional diffusion. Comparison with neutron scattering measurements. Mol Phys 56:1281–1305

    Google Scholar 

  • Warshel A (1984) Dynamics of enzymatic reactions. Proc Natl Acad Sci USA 81:444–448

    Google Scholar 

  • Wien RW, Morrisett JD, McConnell HM (1972) Spin-label-induced nuclear relaxation. Distances between bound saccharides, histidine-15, and tryptophan-123 on lysozyme in solution. Biochemistry 11:3707–3716

    Google Scholar 

  • Witte A (1971) Induktionsanordnung in the ESR-Spektroskopie, Dissertation, Ruhr-Universität, Bochum

    Google Scholar 

  • Yin JJ, Hyde JS (1987) Application of rate equations to ELDOR and saturation recovery experiments on 14N: 15N spin-label pairs. J Magn Res 74:82–93

    Google Scholar 

  • Yin JJ, Pasenkiewicz-Gierula M, Hyde JS (1987) Lateral diffusion of lipids in membranes by pulse saturation recovery electron spin resonance. Proc Natl Acad Sci USA 84:964–968

    Google Scholar 

  • Yin JJ, Feix JB, Hyde JS (1988) Solution of the nitroxide spin-label spectral overlap problem using pulse electron spin resonance. Biophys J 53:525–531

    Google Scholar 

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Authors and Affiliations

  1. Institut für Biophysik, Ruhr-Universität Bochum, Universitätsstrasse, 150, W-4630, Bochum, Germany

    Heinz -Jürgen Steinhoff, Olaf Dombrowsky, Christine Karim & Christoph Schneiderhahn

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  1. Heinz -Jürgen Steinhoff
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  2. Olaf Dombrowsky
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  3. Christine Karim
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  4. Christoph Schneiderhahn
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Steinhoff, H.J., Dombrowsky, O., Karim, C. et al. Two dimensional diffusion of small molecules on protein surfaces: an EPR study of the restricted translational diffusion of protein-bound spin labels. Eur Biophys J 20, 293–303 (1991). https://doi.org/10.1007/BF00450565

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  • DOI: https://doi.org/10.1007/BF00450565

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