Distance-dependent fluorescence quenching of N-acetyl-L-tryptophanamide by acrylamide and iodide

Bogumil Zelent; Ignacy Gryczynski; Jozef Kusba; Michael L. Johnson; Joseph R. Lakowicz

doi:10.1117/12.182750

17 August 1994 Distance-dependent fluorescence quenching of N-acetyl-L-tryptophanamide by acrylamide and iodide

Bogumil Zelent, Ignacy Gryczynski, Jozef Kusba, Michael L. Johnson, Joseph R. Lakowicz

Proceedings Volume 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV; (1994) https://doi.org/10.1117/12.182750
Event: OE/LASE '94, 1994, Los Angeles, CA, United States

Abstract

Frequency-domain fluorometry was used to investigate the time- dependent intensity decays of N-acetyl-L-tryptophanamide (NATA) when collisionally quenched by acrylamide and iodide in propylene glycol at 20 degree(s)C. The intensity decays of NATA became increasingly heterogeneous in the presence of quenching, and the steady-state Stern-Volmer plots show significant upward curvature. These frequency-domain and steady-state data clearly indicate that the rate constants for quenching of NATA by acrylamide and iodide depend exponentially on the fluorophore- quencher separation distance. The NATA intensity decays were not consistent with the Smoluchowski-Collins-Kimball radiation boundary condition for quenching. The upward curvature of the Stern-Volmer plots provide a sensitive method to determine the characteristic distances for fluorophore-quencher interaction. The rate constant for quenching are discussed based on the mechanism of interaction between tryptophanyl fluorophore and quencher molecules which appear to involve electron transfer for acrylamide and heavy atom effects for iodide. These results suggest that the distance-dependent interaction and the effects of solvent dynamics need to be considered in the interpretations of data for quenching of proteins.

Citation Download Citation

Bogumil Zelent, Ignacy Gryczynski, Jozef Kusba, Michael L. Johnson, and Joseph R. Lakowicz "Distance-dependent fluorescence quenching of N-acetyl-L-tryptophanamide by acrylamide and iodide", Proc. SPIE 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV, (17 August 1994); https://doi.org/10.1117/12.182750

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