PMID: 8590598Jul 1, 1995Paper

Fluorescence study of Escherichia coli cyclic AMP receptor protein

Journal of Protein Chemistry
M WasylewskiZ Wasylewski

Abstract

Time-resolved, steady-state fluorescence and fluorescence-detected circular dichroism (FDCD) have been used to resolve the fluorescence contributions of the two tryptophan residues, Trp-13 and Trp-85, in the cyclic AMP receptor protein (CRP). The iodide and acrylamide quenching data show that in CRP one tryptophan residue, Trp-85, is buried within the protein matrix and the other, Trp-13, is moderately exposed on the surface of the protein. Fluorescence-quenching-resolved spectra show that Trp-13 has emission at about 350 nm and contributes 76-83% to the total fluorescence emission. The Trp-85, unquenchable by iodide and acrylamide, has the fluorescence emission at about 337 nm. The time-resolved fluorescence measurements show that Trp-13 has a longer fluorescence decay time. The Trp-85 exhibits a shorter fluorescence decay time. In the CRP-cAMP complex the Trp-85, previously buried in the apoprotein becomes totally exposed to the iodide and acrylamide quenchers. The FDCD spectra indicate that in the CRP-cAMP complex Trp-85 remains in the same environment as in the protein alone. It has been proposed that the binding of cAMP to CRP is accompanied by a hinge reorientation of two protein domains. This allows for penetration of th...Continue Reading

References

Oct 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·J KimS Garges
May 15, 1992·European Journal of Biochemistry·Z WasylewskiK Slizowska
Apr 6, 1990·Biochimica Et Biophysica Acta·B F Luisi, P B Sigler
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Sep 15, 1987·European Journal of Biochemistry·Z Wasylewski, M R Eftink
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Citations

Oct 1, 1996·Journal of Protein Chemistry·P KaszyckiZ Wasylewski
Apr 11, 2009·Proceedings of the National Academy of Sciences of the United States of America·Nataliya PopovychCharalampos G Kalodimos
Feb 10, 2017·Metallomics : Integrated Biometal Science·Aleksandra M BondžićVesna M Vasić
Feb 1, 1997·European Journal of Biochemistry·J Małecki, Z Wasylewski
Jan 30, 2020·Luminescence : the Journal of Biological and Chemical Luminescence·Said Abd El-Monem HassanNadia Fayek Youssef
May 10, 2001·Biochimica Et Biophysica Acta·J G Harman
Apr 16, 2003·Journal of Photochemistry and Photobiology. B, Biology·Roopa Kenoth, Musti J Swamy

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