Abstract
This paper deal with the spectroscopic analysis of proteins, i.e., human serum albumin (HSA) and bovine serum albumin (BSA). These two proteins present very similar UV absorption and fluorescence spectra. The UV-spectra of both proteins result from the sum of the absorption spectra of the aromatic amino acid, mainly tryptophan and tyrosine, present in the biological molecules. On the contrary, the fluorescence HSA and BSA spectra are not the sum of the emission fluorescence spectra of tyrosine and tryptophan, but they are mainly constituted by the tryptophan fluorescence with a poor contribution arising from tyrosine. This fact may be due to inter- and intramolecular quenching phenomena. Significant difference were recorder in the life time fluorescence decay of HSA and BSA, which showed T values of 2.3 and 4.5 nanoseconds, respectively. The time-resolved fluorescence spectroscopy was performed with an experimental apparatus assembled in our laboratories, which is able of measuring the fluorescence decay in the range 2xl0-1010-2xl0-8 sec.
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References
Ballestrieri C., Colonna G., Giovane A., Irace G., and Servillo L., 1978, Amino acid composition and physic-chemical properties of bluefin tuna (Thunnus thynnus) myoglobin, Eur. J. Biochem. 90: 433.
Ballestrieri C., Colonna G., Giovane A., Irace G., and Servillo L., 1980, Second-derivative spectroscopy of proteins: studies on tyrosyl residues, Anal Biochem. 106: 49
Bencze W.L., and Schmid K., 1957, Deth of tyrosine and tryptophan in proteins, Anal. Chem. 29: 1193
Bertini J., Mannucci C., Noferini R., Perico A., and Rovero R, 1993, Rapid simultaneous determination of tryptophan and tyrosine in synthetic peptides by derivative spectroscopy, J. Pharm. Sci. 82:179
Breton J., Martin J.L., Migus A., Antonetti A., and Orsag A., 1986, Femtosecond spectroscopy of excitation energy transfer and initial charge separation in the reaction centre of the photosynthetic bacterium Rhodopseudomonas viridis, Proc. Natl Acad. Sci. USA 83: 5121
Chang J.Y., Knecht R., and Braun D.G., 1983, Isolation and sequence determination of polypeptides at the picomole level, Methods Enzymol. 91: 41
Chen R.F., Vurex G.G., and Alexander N., 1967, Fluorescence decay times: proteins, coenzymes, and other compounds in water, Science 156: 949
Creighton S., Hwang J.K., Warshell A., Parson W.W., and Norris J., 1988, Simulating the dynamics of the primary charge separation process in bacterial photosynthesis, Biochemistry 27: 774
Godik V.I., Blankenship R.E., Causgrove T.P., and Woodbury N., 1993, Time-resolved tryptophan fluorescence in photosynthetic reaction centres from Rhodobacter sphaeroides, FEBS Letters 321: 229.
Goodwin T.W., and Morton R.A., 1946, Deth of carotene and vitamin A in butter and margarine, Biochem. J. 40: 628.
Hassan S.S.M., 1975, New spectrophotometric method for simultaneous determination of tryptophan and tyrosine, Anal Chem. 47: 1429
Hochstrasser R.M. and Johnson C.K., 1988, Biological Processes Studied by Applied Physics, in: Ultrashort Laser Pulses and Application, W. Kaiser, Ed., vol. 60, Springer-Verlag, Berlin, p. 357
Horton H.R., and Koshland D.E., 1965, Highly reactive coloured reagent with selectivity for the tryptophan residue in proteins -2-hydroxy-5-nitrobenzyl bromide, J. Am. Chem. Soc. 87: 1126
Jones B.N., Paabo S., and Stein S., 1981, Amino acid analysis and enzymatic sequence determination of peptides by an improved o-phtaldialdehyde percolum labelling procedure, Liq. Chromatogr. 4: 565
Mantsch H.H., Moffatt D.J., and Casal H.L., 1988, Fourier transform methods for spectral resolution enhancement, J. Mol. Struct. 173: 285
Mattioli T.A, Gray K.A., Lutz M., Oesterhelt D., and Bruno R., 1991, Resonance raman characterization of Rhodobacter sphaeroides reaction centres bearing site-directed mutations at tyrosine M210, Biochemistry 30: 1715.
Molnär-Perl I., and Pinter-Szakacs M., 1989, Spectrophotometric determination of tryptophan in intact proteins by the acid ninhydrin method, Anal Biochem. 177: 16
Parson W.W., Chu Z., and Warshell A., 1990, Microscopic simulation of quantum dynamics and nuclear funnelling in bacterial reaction centres, Biochim. Biophys. Acta 1017: 251
Servillo L., Colonna G., Ballestrieri C., Ragone R., and Irace G., 1982, Simultaneous determination of tyrosine and tryptophan residues in proteins by second-derivative spectroscopy, Anal Biochem. 126: 251
Simpson R.J., Moritz R.L., Nice E.C., Grego B., Yoshizaki F., Sugimura Y., Freeman H.C., and Murata M., 1986, Complete amino acid sequence of plastocyanin from a green alga, Enteromorphaprolifera, Eur. J. Biochem. 157: 497.
Spies J.R., 1967, Allergeus (XIX) no. of antigens and the homogeneity of the isolated antigens of fraction CB-lAfrom castor beans, Anal Chem. 39: 1412.
Teutlein H., Shulten K., Deisenhofer J., Michel H., Brunger A., and Karplus M., 1992, in: The Photosynthetic Bacterial Reaction Centre, Structure and Dynamics, NATO ASI Series A 149, J. Breton and A. Vermeglio, Eds., Plenum Press, New York, p. 139
Villari A., Micali N., Fresta M., and Puglisi G., 1992, Simultaneous spectrophotometric determination in solid phase of aspirin and its impurity salicylic acid in pharmaceutical formulations, J. Pharm. Sci. 81: 895
Villari A., Micali N., Fresta M., and Puglisi G., 1994, Spectrofluorimetry at zero angle: determination of salicylic acid in acetylsalicylic acid pharmaceutical formulations, The Analyst 119: 1561
Woodbury N.W., Becker M., Middendorf P., and Parson W.W., 1985, Nanosecond fluorescence from isolated photosynthetic reaction centres of Rhodopseudomonas sphaeroides, Biochemistry 24: 7516
Wuite A., 1959, Effect of pH on fluorescence of tyrosine, tryptophan and related compounds, Biochem. J. 71: 217.
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© 1996 Plenum Press, New York
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Villari, A., Micali, N., Fresta, M., Trusso, S., Puglisi, G. (1996). Time-Resolved Fluorescence. In: Filippini, G.A., Costa, C.V.L., Bertazzo, A. (eds) Recent Advances in Tryptophan Research. Advances in Experimental Medicine and Biology, vol 398. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-0381-7_117
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DOI: https://doi.org/10.1007/978-1-4613-0381-7_117
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