Probing iso-1-cytochrome c structure by site-directed spin labeling and electron paramagnetic resonance techniques

• Autorzy: Pyka J. , Osyczka A. , Turyna B. , Blicharski W. , Froncisz W.
• Języki publikacji: EN

Abstrakty

EN

A cysteine-specific methanethiosulfonate spin label was introduced into yeast iso-1-cytochrome c at three different positions. The modified forms of cytochrome c included: the wild-type protein labeled at naturally occurring C102, and two mutated proteins, S47C and L85C, labeled at positions 47 and 85, respectively (both S47C and L85C derived from the protein in which C102 had been replaced by threonine). All three spin-labeled protein derivatives were characterized using electron paramagnetic resonance (EPR) techniques. The continuous wave (CW) EPR spectrum of spin label attached to L85C differed from those recorded for spin label attached to C102 or S47C, indicating that spin label at position 85 was more immobilized and exhibited more complex tumbling than spin label at two other positions. The temperature dependence of the CW EPR spectra and CW EPR power saturation revealed further differences of spin-labeled L85C. The results were discussed in terms of application of the site-directed spin labeling technique in probing the local dynamic structure of iso-1-cytochrome c.

Słowa kluczowe

EN

cytochrome bc1 complex; yeast; cytochrome c; iso-1-cytochrome c; electron paramagnetic resonance; Saccharomyces cerevisiae

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1999
• Tom: 46
• Numer: 4
• Opis fizyczny: p.889-899,fig.

Twórcy

autor

Pyka J.

• Jagiellonian University, Al.A.Mickiewicza 3, 31-120 Krakow, Poland

autor

Osyczka A.

autor

Turyna B.

autor

Blicharski W.

autor

Froncisz W.

Bibliografia

• 1. Bushnell, G.W., Louie, G.V. & Brayer, G.D. (1990) High-resolution three-dimensional structure of horse heart cytochrome c. J. Mol Biol. 214, 585-595.
• 2. Ochi, li., Hata, Y., Tanaka, N., Kakudo, M., Sakurai, T., Aihara, S. & Morita, Y. (1983) Structure of rice ferricytochrome c at 2.0 A resolution. J. Mol Biol 166. 407-418.
• 3. Takano, T., Trus, B.L., Mandel, N., Mandel, G., Kallai, O.B., Swanson, R. & Dickerson, R.E. (1977) Tuna cytochrome c at 2.0 Â resolu­tion. II. Ferrocytochrome structure analysis. J. Biol Chem. 252, 776-785.
• 4. Louie, G.V. & Brayer, G.D. (1990) High-resolu­tion refinement of yeast iso-l-cytochrome c and comparisons with other eukaryotic cytochromes c. J. Mol Biol 214, 527-555.
• 5. Berghuis, A.M. & Brayer, G.D. (1992) Oxida­tion state-dependent conformational changes in cytochrome c. J. Mol Biol 223, 959-976.
• 6. Berghuis, A.M., Guillemette, J.G., McLendon, G., Sherman, F., Smith, M. & Brayer, G.D. (1994) The role of conserved internal water molecule and its associated hydrogen bond network in cytochrome c. J. Mol Biol 236, 786-799.
• 7. Baistrocchi, P., Banci, L., Bertini, I., Turano, P., Bren. K.L. & Gray, H.B. (1996) Three-di­mensional solution structure of Saccha- romyces cereuisiae reduced iso-l-cytochrome c. Biochemistry 35, 13788-13796.
• 8. Banci, L., Gori-Savellini, G. & Turano, P. (1997) A molecular dynamics study in explicit water of the reduced and oxidized forms of yeast iso-l-cytochrome c. Solvation and dy­namic properties of the two oxidation states. Eur. J. Biochem. 249, 716-723.
• 9. Banci. L., Bertini, I., Bren, K.L., Gray, H.B., Sompornpisut, P. & Turano, P. (1997) Solu­tion structure of oxidized Saccharomyces cerevisiae iso-l-cytochrome c. Biochemistry 36, 8992-9001.
• 10. Margoliash, E. & Schejter, A. (1996) How does a small protein become so popular?: A succinct account of the development of our understand­ing of cytochrome c, in Cytochrome c: a Multidisciplinary Approach (Scott, R.A. & Mauk, A.G., eds.) pp. 3-31, University Science Books, Sauaalito, California.
• 11. Moore, G.R. & Pettigrew, G.W. (1987) Cyto­chromes c: Evolutionary, Structural and Phy~ sicochemical Aspects, pp. 27-107, Springer- Verlag, Berlin.
• 12. Hampsey, D.M., Das, G. & Sherman, F. (1986) Amino acid replacement in yeast iso-l-cyto­chrome c. J. Biol Chem. 261, 3259-3271.
• 13. Caffrey, M.S. & Cusanovich, M.A. (1994) Site-specific mutagenesis studies of cytochro­mes c. Biochim. Biophys. Acta 1187,277-288.
• 14. Hubbell, W.L., Mchaourab, H.S., Altenbach, C. & Lietzow, M.A. (1996) Watching proteins move using sit^directed spin labeling. Struc­ture 4, 779-783.
• 15. Shin, Y., Levinthal, C., Levinthal, F. & Hubbell, W.L. (1993) Colicin El binding to membranes: Time-resolved studies of spin-labeled mutants. Science 259, 960-963.
• 16. Altenbach, C., Flitsch, S.L, Khorana, H.G. & Hubbell, W.L. (1989) Structural studies on transmembrane proteins: 2. Spin labeling of bacteriorhodopsin mutants at unique cyste­ines. Biochemistry 28, 7806-7812.
• 17. Altenbach, C., Greenhalgh, D.A., Khorana, H.G. & Hubbell, W.L. (1994) A collision gradi­ent method to determine the immersion depth of nitroxides in lipid bilayers: Application to spin-labeled mutants of bacteriorodopsin. Proc. NatL Acad. Sci. U.S.A. 91, 1667-1671.
• 18. Altenbach, C., Yang, K., Farrens, D.L., Farahbakhsh, Z.T., Khorana, H.G. & Hubbell, W.L. (1996) Structural features and light-dependent changcs in the cytoplasmic interhelical E-F loop region of rhodopsin: A site-directed spin-labeling study. Biochemis­try 35,12470-12478.
• 19. Mchaourab, H.S., Lietzow, M.A., Hideg, K. & Hubbell, W.L. (1996) Motion of spin-labeled side chains in T4 lysozyme. Correlation with protein structure and dynamics. Biochemistry 35, 7692-7704.
• 20. Oh, K.J., Zhan, H.f Cui, C., Hideg, K., Collier, R.J. & Hubbell, W.L. (1996) Organization of diphtheria toxin T domain in bilayers: a site-directed spin labeling study. Science 273, 810-812.
• 21. Voss, J.. He, M., Hubbell, W.L. & Kaback, H.R. (1996) Site-directed spin labeling demon­strates that transmembrane domain XII in the lactose permease of Escherichia coli is an al­pha-helix. Biochemistry 35, 12915-12918.
• 22. Lindgren, M., Svensson, M., Freskg^rd, P.O., Carlsson, U., Jonasson, P., M&rtensson, L.G. & Jonsson, B.H. (1995) Characterization of a folding intermediate of human carbonic anhydrase II: Probing local mobility by elec­tron paramagnetic resonance. Biophys. J. 69, 202-213.
• 23. Moench, S.J. & Satterlee, J.D. (1995) A com­parison of spectral and physicochemical prop­erties of yeast iso-l-cytochrome c and Cys 102-modified derivatives of the protein. J. Pro­tein Chem. 14, 567-582.
• 24. Drott, H.R., Lee, C.P. & Yonetani, T. (1970) Spin label studies of hemoproteins: I. Cyto­chrome c. J. Biol Chem. 245, 5875-5879.
• 25. Vanderkooi, J., Erecińska, M. & Chance. B. (1973) Cytochrome c interaction with mem­branes. II. Comparative study of the interac­tion of c cytochromes with the mitochondrial membrane. Arch. Biochem. Biophys. 157, 531-540.
• 26.Snel, M.M.E., de Kruijff, B. & Marsh. D. (1994) Interaction of spin-labeled apocyto- chrome c and spin-labeled cytochrome c with negatively charged lipids studied by electron spin resonance. Biochemistry 33, 7146-7156.
• 27. Snel, M.M.E., de Kruijff, B. & Marsh, D. (1994) Membrane location of spin-labeled apocytochrome c and cytochrome c deter­mined by paramagnetic relaxation agents. Bio­chemistry 33,11150-11157.
• 28. Qu, K., Vaughn, J.L., Sienkiewicz, A, Scholes, C.P. & Fetrow, J.S. (1997) Kinetics and motional dynamics of spin-labeled yeast iso-l-cytochrome c: 1. Stopped-flow electron paramagnetic resonance as a probe for protein folding/unfolding of the C-terminal helix spin-labeled at cysteine 102. Biochemistry 36, 2884-2897.
• 29. Brayer, G.D. & Murphy, M.E.P. (1996) Struc­tural studies of eukaryotic cytochromes c; in Cytochrome c: A Multidisciplinaty Approach (Scott, R.A. & Mauk, A.G., eds.) pp. 103-166, University Science Books, Sausalito, Califor­nia.
• 30. Cutler, R.L., Pielak, G.J., Mauk, A.G. & Smith, M. (1987) Replacement of cysteine-107 of Saccharomyces cerevisiae iso-l-cytochrome c with threonine: Improved stability of the mu­tant protein. Protein Eng. 1, 95-99.
• 31. Faye, G., Leung, D.W., TatcheU, K., Hall, B.D. & Smith, M. (1981) Deletion mapping of se­quences essential for in vivo transcription of the iso-l-cytochrome c gene. /Yoc. Natl. Acad. Sci. U.S.A. 78, 2258-2262.
• 32. Sherman, F. (1991) Getting started with yeast. Methods EnzymoL 194, 3-21.
• 33.Inglis, S.C., Guillemette, J.G., Johnson, J. A. & Smith, M. (1991) Analysis of the invariant Phe82 residue of yeastrl-cytochrome c by site-directed mutagenesis using a phagemid yeast shuttle vector. I*rotein Eng. 4,569-574.
• 34. Horton, R.M.. Cai, Z.L., Ho, S.N. & Pease, L.R. (1990) Gene splicing by overlap extension: Tailor-made genes using the polymerase chain reaction. Bioiechniques 8, 528-535.
• 35.Ito, H., Fukuda, Y., Murata, K. & Kimura, A. (1983) Transformation of intact yeast cells treated with alkali cations. J. Bacteriol. 153, 163-168.
• 36.Soni, R., Carmichael, J.P. & Murray, J.A. (1993) Parameters affecting lithium ace­tate-mediated transformation of Saccharo­myces cerevisiae and development of a rapid and simplified procedure. Curr. Genet. 24, 455-459.
• 37. Pielak, G.J., Mauk, A.G. & Smith, M. (1985) Site-directet mutagenesis of cytochrome c shows that an invariant Phe is not essential for function. Nature 313, 152-154.
• 38. Bowler, B.E., May, K., Zaragoza, T., York, P., Dong, A. & Coughey, W.S. (1993) Desta­bilizing effects of replacing a surface lysine of cytochrome c with aromatic amino acids: Implications for the denatured state. Biochem­istry Z2, 183-190.
• 39. Turyna, B., Osyczka, A., Kostrzewa, A., Blicharski, W., Enghild, J.J. & Froncisz, W. (1998) Preparation and electron paramagnetic resonance characterization of spin labeled monoderivatives of horse cytochrome c. Biochim. Biophys. Acta 1386, 50-58.
• 40. Lo, T.P., Guillemette, J.G., Louie, G.V., Smith, M. & Brayer, G.D. (1995) Structural studies of the roles of residues 82 and 85 at the interac­tive face of cytochrome c. Biochemistry 34, 163-171.
• 41. Kraulis, P. (1991) MOLSCRIPT: A program to produce both detailed and schemate plots of proteins. J. Appl. Cryst. 24, 946-950.
• 42. Merritt, E.A. & Murphy, M.E.P (1994) Raster3D version 2.0: A program for photo­realistic molecular graphics. Acta Crystallogr. Sect. D 50, 869-873.