Time-resolved and steady-state fluorescence quenching of N-acetyl-L-tryptophanamide by acrylamide and iodide.
We examined the time-resolved and steady-state fluorescence quenching of N-acetyl-L-tryptophanamide (NATA) by acrylamide and iodide, over a range of viscosities in propylene glycol. The quenching of NATA by acrylamide and iodide results in heterogeneity of the intensity decay which increases with the quencher concentration. We attribute the complex decays of NATA to transient effects in diffusion and the nature of the fluorophore-quencher interaction. These data were compared using the phenomenological radiation boundary condition (RBC) and distance-dependent quenching (DDQ) models for collisional quenching. We used global analysis of the time-resolved frequency-domain and steady-state data to select between the models. Consideration of both the frequency-domain and steady state data demonstrate that the quenching rate depends exponentially on the fluorophore-quencher distance, indicating the validity of the DDQ model. The rate constants for acrylamide and iodide quenching, at the constant distance of 5 A, were found to be near 10(13) s-1 and 10(9) s-1, respectively. These rates reflect electron transfer and exchange interactions as the probable quenching mechanisms, respectively.
Department of Biochemistry and Molecular Biology, University of Maryland, Baltimore School of Medicine 21201, USA.
B
B
Zelent
J
J
Kuśba
I
I
Gryczynski
M L
ML
Johnson
J R
JR
Lakowicz
D020106:Acrylamide; D000178:Acrylamides / Q000737:chemistry*; D005453:Fluorescence; D008432:Mathematical Computing; D019946:Propylene Glycol; D011506:Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014364:Tryptophan / Q000031:analogs & derivatives* / Q000737:chemistry; D014783:Viscosity
D000178:Acrylamides; D011506:Proteins; D020106:Acrylamide; C016952:N-acetyltryptophanamide; D019946:Propylene Glycol; D014364:Tryptophan