Abstract
With few exceptions, the goal of most protein purification efforts is to obtain a sample that is not only pure, but that also maintains the protein in its native (i.e., biologically active) conformation. The ability to describe the conformation of a protein in solution, and to relate changes in conformation with biological activity, is thus a major focus of protein science. The most detailed description of protein structures come from the determination of the complete three-dimensional arrangement of protein components in space, from x-ray crystallographic or nuclear magnetic resonance (NMR) studies. Despite their power, however, these methods are not without their attendant drawbacks. X-ray diffraction studies of proteins are dependent on obtaining protein crystals of sufficient size and quality to yield usable diffraction patterns. This can often be a time consuming, and not necessarily successful, undertaking. Even when high quality crystals are obtained, solving the structure from the resulting diffraction patterns is a laborious and time consuming effort. Add to this the fact that certain classes of proteins, such as integral membrane proteins, are inherently difficult to crystallize, and one soon realizes that x-ray crystallography, while an extremely powerful method, is not a panacea for protein structural problems. Multidimensional NMR spectroscopy likewise suffers from certain difficulties that restrict its utility. Perhaps the greatest limit to the use of NMR spectroscopy for solving protein structures is that the complexity of the multidimensional data is such that the size of a protein that can reasonably be solved is limited to about 100 amino acids or so. While significant efforts are currently being put forth to push up the size limit for NMR spectroscopy, at least for the present this method is limited to relatively small proteins.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Campbell, I. D., and Dwek, R. A. (1984) Biological Spectroscopy, Benjamin/Cummings. Menlo Park, CA.
Cantor, C. R., and Schimmel, P. R. (1980) Biophysical Chemistry, Part II, W. H. Freeman, San Francisco, CA.
Chang, C. T.; Wu, C.-S. C.; and Yang, J. T. (1978) Analyt. Biochem., 91, 12.
Chen, R. F., and Edelhoch, H. (1975) Biochemical Fluorescence, Vols. 1 and 2, Marcel Dekker, New York.
Chen, Y. H., and Yang, J. T. (1971) Biochem. Biophys. Res. Commun., 44, 1285.
Copeland, R. A.; Ji, H.; Halfpenny, A. J.; Williams, R. W.; Thompson, K. C.; Herber, W. K.; Thomas, K. A.; Brunner, M. W.; Sitrin, R. D.; Yamazaki, S.; and Middaugh, C. R. (1991) Arch. Biochem. Biophys., 289, 53–61.
Demchenko, A. P. (1986) Ultraviolet Spectroscopy of Proteins, Springer-Verlag, New York.
Donovan, J. W. (1973) Meth. Enzymol., 27, 497–525.
Fodor, S. P. A.; Copeland, R. A.; Grygon, C. A.; and Spiro, T. G. (1989) J. Am. Chem. Soc. USA, 111, 5509.
Freifelder, D. (1982) Physical Biochemistry, W. H. Freeman, San Francisco, CA. Herskovits, T. T., and Sorensen, M. (1968) Biochemistry, 7, 2533–2542. Holzwarth, G. N., and Doty, P. (1965) J. Am. Chem. Soc., 87, 218.
Kotake, S.; Hey, P.; Mirmira, R. G.; and Copeland, R. A. (1991) Arch. Biochem. Biophys., 285, 126–133.
Lackowicz, J. R. (1983) Principles of Fluorescence Spectroscopy,Plenum Press, New York.
Lehrer, S. S. (1971) Biochemistry, 10, 3254–3263.
Parker, F. S. (1983) Applications of Infrared, Raman, and Resonance Raman Spectroscopy in Biochemistry, Plenum Press, New York.
Ragone, R.; Colonna, G.; Balestrieri, C.; Servillo, L.; and Irace, G. (1984) Biochemistry, 23, 1871–1875.
Yang, J. T.; Wu, C.-S. C.; and Martinez, H. M. (1986) Meth. Enzymol.,130 208269.
Zabin, H. B. (1991) Ph.D. Thesis, The University of Chicago, Chicago, Illinois. Zabin, H. B., and Terwilliger, T. C. (1991) J. Mol. Biol., 219, 257–275.
Author information
Authors and Affiliations
Rights and permissions
Copyright information
© 1994 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Copeland, R.A. (1994). Spectroscopic Probes of Protein Structure. In: Methods for Protein Analysis. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-1505-7_9
Download citation
DOI: https://doi.org/10.1007/978-1-4757-1505-7_9
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4757-1507-1
Online ISBN: 978-1-4757-1505-7
eBook Packages: Springer Book Archive