PMID: 2502172May 2, 1989Paper

Confirmation that multiexponential fluorescence decay behavior of holoazurin originates from conformational heterogeneity

Biochemistry
C M Hutnik, A G Szabo

Abstract

Homologous azurins from Pseudomonas fluorescens (ATCC 13525) and Pseudomonas aeruginosa (ATCC 10145) were examined by a number of electrophoretic techniques, and their copper to protein stoichiometry was determined by atomic absorption and amino acid analysis. Provided that the spectral ratio (A620/A280 or A625/A280) was 0.53 and there was no evidence of a Soret band in the absorption spectrum, then these criteria can be used to judge the homogeneity of the azurin sample. If the spectral ratio was less than 0.50, evidence suggested a nonreconstitutable, non-trypsin-digestible apoazurin was present. The fluorescence decay of these homogeneous holoazurins included three components, not two as previously reported [Szabo, A. G., et al. (1983) Biophys. J. 41, 233-244]. Whereas the decay times were nearly the same for the azurins from the different sources, the fractional fluorescence of each component varied with the azurin measured. The fluorescence of the corresponding apoazurins, prepared by a refined procedure, obeyed monoexponential decay kinetics. The temperature and pH effects on the fluorescence behavior of these homologous azurins are presented with the pH study suggesting an influence by a group which titrates between pH 5...Continue Reading

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Citations

May 1, 1993·Protein Science : a Publication of the Protein Society·W M AtkinsJ J Villafranca
Jan 1, 1990·Biology of Metals·N RosatoA Finazzi Agrò
Aug 7, 1993·Biochimica Et Biophysica Acta·W S OstrowskiK Yagi
Oct 3, 1999·European Journal of Biochemistry·G MeiA Finazzi-Agrò
Oct 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·D Ehrenstein, G U Nienhaus
Sep 22, 2001·Protein Science : a Publication of the Protein Society·A Ababou, E Bombarda
Jan 11, 2001·Protein Science : a Publication of the Protein Society·B UllrichJ Fidy
Jul 15, 1995·European Journal of Biochemistry·J SalgadoJ M Moratal
Aug 7, 1990·Biochemistry·J E HansenG R Fleming
Jan 1, 1991·Archives of Biochemistry and Biophysics·N RosatoG W Canters
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Mar 15, 1995·European Journal of Biochemistry·J M MoratalH R Jiménez
Apr 20, 2004·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·P P PompaG W Canters

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