- Access by Xinjiang University
Structural stability study of protein monolayers in air
Phys. Rev. E 69, 032901 – Published 30 March, 2004
DOI: https://doi.org/10.1103/PhysRevE.69.032901
Abstract
The assessment of the folding and of the structural stability of a protein in air, upon immobilization in the solid state, represents a critical point from both a fundamental point of view and for the development of solid state nanobioelectronics. The recent demonstrations by Rinaldi et al. [R. Rinaldi et al., Adv. Mater. 14, 1453 (2002); Appl. Phys. Lett. 82, 472 (2003); Ann. (N.Y.) Acad. Sci. 1006, 187 (2003)] of protein-based solid state devices and transistors working in air have raised an intriguing question about the behavior of a biomolecule under nonphysiological conditions. The operation principle of the realized devices is based on the physiological electron transfer function of the metalloprotein azurin. This means that azurin should retain its shape and functionality also in the solid state when utilized in air and at room temperature. In this Brief Report, we prove this claim by analyzing the conformational state of the azurin monolayers developed for such devices by means of intrinsic fluorescence spectroscopy. We show that the immobilization of azurins in the solid state under nonliquid conditions, by means of a specific chemisorption process, does not necessarily lead to protein denaturation. This result is of great importance because it opens up interesting perspectives for the development of solid state hybrid nanodevices for electronic applications requiring nonliquid environments.
References (29)
- N.K. Chaki and K. Vijayamohanan, Biosens. Bioelectron. 17, 1 (2002).
- E.T. Adman, in Topics in Molecular and Structural Biology: Metalloproteins, edited by P.M. Harrison (Chemie Verlag, Weinheim, 1985), p. 173.
- E.T. Adman, Adv. Protein Chem. 42, 144 (1991).
- A.G. Sykes, Adv. Inorg. Chem. 36, 377 (1991).
- L. Andolfi, S. Cannistraro, G.W. Canters, P. Facci, A.G. Ficca, I.M.C. Van Amsterdam, and M.P. Verbeet, Arch. Biochem. Biophys. 339, 81 (2002).
- P. Facci, D. Alliata, and S. Cannistraro, Ultramicroscopy 89, 291 (2001).
- P. Facci, D. Alliata, L. Andolfi, B. Shnyder, and R. Koetz, Surf. Sci. 504, 282 (2002).
- A.K. Gaigalas and G. Niaura, J. Colloid Interface Sci. 193, 60 (1997).
- J.J. Davis, C.M. Halliwell, A.H.O. Hill, G.W. Canters, M.C. Van Amsterdam, and M.P. Verbeet, New J. Chem. 1, 1119 (1998).
- E.P. Friis, J.E. Andersen, Y.I. Kharkats, A.M. Kuznetsov, R.J. Nichols, J.D. Zhang, and J. Ulstrup, Proc. Natl. Acad. Sci. U.S.A. 96, 1379 (1999).
- Q. Chi, J. Zhang, J.U. Nielsen, E.P. Friis, I. Chorkendorff, G.W. Canters, J.E.T. Andersen, and J. Ulstrup, J. Am. Chem. Soc. 122, 4047 (2000).
- R. Rinaldi, A. Biasco, G. Maruccio, R. Cingolani, D. Alliata, L. Andolfi, P. Facci, F. De Rienzo, R. Di Felice, and E. Molinari, Adv. Mater. (Weinheim, Ger.) 14, 1453 (2002).
- R. Rinaldi, A. Biasco, G. Maruccio, V. Arima, P. Visconti, R. Cingolani, P. Facci, F. De Rienzo, R. Di Felice, E. Molinari, M.P. Verbeet, and G.W. Canters, Appl. Phys. Lett. 82, 472 (2003).
- R. Rinaldi, G. Maruccio, A. Biasco, P. Visconti, S. D’Amico, and R. Cingolani, Ann. (N.Y.) Acad. Sci. 1006, 187 (2003).
- A. Finazzi-Agrò, G. Rotilio, L. Avigliano, P. Guerrieri, V. Boffi, and B. Mondovì, Biochemistry 9, 2009 (1970).
- H. Nar, A. Messerschmidt, R. Huber, M. Van de Kamp, and G.W. Canters, J. Mol. Biol. 221, 765 (1991).
- S.J. Kroes, G.W. Canters, G. Gilardi, A. Van Hoek, and A.J. Visser, Biophys. J. 75, 2441 (1998).
- E.N. Baker, J. Mol. Biol. 203, 1071 (1988).
- H. Nar, A. Messerschmidt, R. Huber, M. Van de Kamp, and G.W. Canters, FEBS Lett. 306, 119 (1992).
- H. Nar, R. Huber, A. Messerschmidt, A.C. Filippou, M. Barth, M. Jaquinod, M. Van de Kamp, and G.W. Canters, Eur. J. Biochem. 205, 1123 (1992).
- A.G. Szabo, T.M. Stepanik, D.M. Wayner, and N.M. Young, Biophys. J. 41, 233 (1983).
- J.W. Petrich, J.W. Longworth, and G.R. Fleming, Biochemistry 26, 2711 (1987).
- C.M. Hutnik and A.G. Szabo, Biochemistry 28, 3923 (1989).
- C.M. Hutnik and A.G. Szabo, Biochemistry 28, 3935 (1989).
- J.E. Hansen, J.W. Longworth, and G.R. Fleming, Biochemistry 29, 7329 (1990).
- J. Leckner, N. Bonander, P. Wittung-Stafshede, B.G. Malmstrom, and B.G. Karlsson, Biochim. Biophys. Acta 1342, 19 (1997).
- J. Leckner, P. Wittung-Stafshede, N. Bonander, B.G. Karlsson, and B.G. Malmstrom, J. Biol. Inorg. Chem. 2, 368 (1997).
- A. Grinvald, J. Schlessinger, I. Pecht, and I.Z. Steinberg, Biochemistry 14, 1921 (1975).
- The reported investigation on the fluorescence energy transfer, however, deserves further comment. We know that in recent years the issue of the Tyr to Trp energy transfer for the azurins has been the subject of debate. Nevertheless, it is beyond the scope of this work to characterize such a process or to measure its specific parameters, but to use the possible dependence of the emission spectrum on the excitation wavelength as a probe to monitor the induction of conformational transitions in the protein. Besides the properties of the Tyr residues and the specific controversies about the energy transfer mechanism, it is a fact that the photoluminescence of azurin is independent of the excitation wavelength if the protein is in the folded state, while it can be strongly affected by the excitation wavelength if the protein is denatured. Hence the wavelength independence observed in the fluorescence emission of the azurin monolayers indeed indicates the existence of proteins which retain the nativelike conformation.