skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Tryptophan and ATTO 590: Mutual Fluorescence Quenching and Exciplex Formation

Abstract

Investigation of fluorescence quenching of probes, such as ATTO dyes, is becoming an increasingly important topic owing to the use of these dyes in super-resolution microscopies and in single-molecule studies. Photoinduced electron transfer is their most important nonradiative pathway. Because of the increasing frequency of the use of ATTO and related dyes to investigate biological systems, studies are presented for inter- and intramolecular quenching of ATTO 590 with tryptophan. In order to examine intramolecular quenching, an ATTO 590–tryptophan conjugate was synthesized. It was determined that tryptophan is efficiently quenching ATTO 590 fluorescence by excited-state charge transfer and two charge transfer complexes are forming. In addition, it was discovered that an exciplex (whose lifetime is 5.6 ns) can be formed between tryptophan and ATTO 590, and it is suggested that the possibility of such exciplex formation should be taken into account when protein fluorescence is monitored in a system tagged with ATTO dyes.

Authors:
 [1];  [1];  [2];  [2];  [1]
  1. Ames Laboratory
  2. Iowa State University
Publication Date:
Research Org.:
Ames Laboratory (AMES), Ames, IA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1163901
Report Number(s):
IS-J 8411
Journal ID: ISSN 1520-6106
DOE Contract Number:
DE-AC02-07CH11358
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Physical Chemistry. B, Condensed Matter, Materials, Surfaces, Interfaces and Biophysical Chemistry; Journal Volume: 118; Journal Issue: 29
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE; Biochemical Methods

Citation Formats

Bhattacharjee, Ujjal, Beck, Christie, Winter, Arthur, Wells, Carson, and Petrich, Jacob W. Tryptophan and ATTO 590: Mutual Fluorescence Quenching and Exciplex Formation. United States: N. p., 2014. Web. doi:10.1021/jp412045m.
Bhattacharjee, Ujjal, Beck, Christie, Winter, Arthur, Wells, Carson, & Petrich, Jacob W. Tryptophan and ATTO 590: Mutual Fluorescence Quenching and Exciplex Formation. United States. doi:10.1021/jp412045m.
Bhattacharjee, Ujjal, Beck, Christie, Winter, Arthur, Wells, Carson, and Petrich, Jacob W. Thu . "Tryptophan and ATTO 590: Mutual Fluorescence Quenching and Exciplex Formation". United States. doi:10.1021/jp412045m.
@article{osti_1163901,
title = {Tryptophan and ATTO 590: Mutual Fluorescence Quenching and Exciplex Formation},
author = {Bhattacharjee, Ujjal and Beck, Christie and Winter, Arthur and Wells, Carson and Petrich, Jacob W},
abstractNote = {Investigation of fluorescence quenching of probes, such as ATTO dyes, is becoming an increasingly important topic owing to the use of these dyes in super-resolution microscopies and in single-molecule studies. Photoinduced electron transfer is their most important nonradiative pathway. Because of the increasing frequency of the use of ATTO and related dyes to investigate biological systems, studies are presented for inter- and intramolecular quenching of ATTO 590 with tryptophan. In order to examine intramolecular quenching, an ATTO 590–tryptophan conjugate was synthesized. It was determined that tryptophan is efficiently quenching ATTO 590 fluorescence by excited-state charge transfer and two charge transfer complexes are forming. In addition, it was discovered that an exciplex (whose lifetime is 5.6 ns) can be formed between tryptophan and ATTO 590, and it is suggested that the possibility of such exciplex formation should be taken into account when protein fluorescence is monitored in a system tagged with ATTO dyes.},
doi = {10.1021/jp412045m},
journal = {Journal of Physical Chemistry. B, Condensed Matter, Materials, Surfaces, Interfaces and Biophysical Chemistry},
number = 29,
volume = 118,
place = {United States},
year = {Thu Jul 24 00:00:00 EDT 2014},
month = {Thu Jul 24 00:00:00 EDT 2014}
}
  • The fluorescence quenching of tyrptophan (Trp) in the absence and presence of 18-crown-6 in CH/sub 3/OH-H/sub 2/ (9:1), vv) mixtures has been studied by means of nanosecond time-resolved single-photon counting, fluorimetry, and photochemical H-D isotope exchange. The fluorescence intensity increases markedly with increasing concentration of 18-crown-6. The quenching can be estimated from the equation k/sub q/ = tau/sub 1//sup 1 -/ - (tau /sub 2//sup max/)/sup -1/, where tau/sub 1/ and tau/sub 1//sup max/ denote the fluorescence lifetimes for free Trp and the 1:1 Trp-18-crown-6 complex, respectively. The internal quenching originates from the electrophilic protonation of the /sup +/NH/sub 3/more » group of Trp at the C-4 position of the excited indole ring plus the charge-transfer interaction between the excited indole ring and the ammonium group. The stabilization constant K/sub g/ for the 1:1 complex of Trp with 18-crown-6 has been determined by means of fluorimetry.« less
  • Using an emission-absorption laser photolysis method, the quenching mechanism of the pyrene-N,N-dimethylaniline exciplex fluorscence by inverted micelles is studied. The rate of enhanced intersystem crossing depends upon water pool size and is reduced by external magnetic fields. 15 refs., 3 figs., 2 tabs.
  • Triplet acetone interacts with mesitylene or durene (in solution at 20 {degrees}C) to generate new triplet transients. These transients were followed by time-resolved single-photon counting, using the fluorescence of 9,10-dibromoanthracene as a triplet probe in acetonitrile and in five aliphatic hydrocarbons. Based on kinetic considerations alone, the new transient could be either a triplet exciplex or the triplet of the methylbenzene, excited by energy transfer from triplet acetone. Solvent polarity has no effect on the rate of transient formation, which varies by a factor of {approx_equal}2 from acetonitrile to cyclohexane, is 2-3 orders of magnitude below the diffusion controlled limit,more » and correlates only with the polarizability of the solvent. This result and the fact that the lifetimes of the transients are compatible with that of the triplet methylbenzenes favor the energy-transfer interpretation of the data. On the other hand, earlier observations with xylene and benzene, where triplet energy transfer would be significantly endothermic, point toward the exciplex interpretation. Attempts at comparing the absorption spectrum of triplet durene with the spectrum previously assigned to the triplet exciplex of durene and acetone{sup 2d} have so far been inconclusive. Definitive proof of quenching via exciplex or via energy transfer is therefore not available now. Both models are discussed. 26 refs., 9 figs., 3 tabs.« less