Inter- and intramolecular fluorescence quenching of organic dyes by tryptophan

被引:262
作者
Marmé, N
Knemeyer, JP
Sauer, M
Wolfrum, J
机构
[1] Univ Bielefeld, Fak Phys, Angewandte Laserphys & Spektroskopie, D-33615 Bielefeld, Germany
[2] Heidelberg Univ, Inst Phys Chem, D-69120 Heidelberg, Germany
关键词
D O I
10.1021/bc0341324
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Steady-state and time-resolved fluorescence measurements were performed to elucidate the fluorescence quenching of oxazine, rhodamine, carbocyanine, and bora-diaza-indacene dyes by amino acids. Among the natural amino acids, tryptophan exhibits the most pronounced quenching efficiency. Especially, the red-absorbing dyes ATTO 655, ATTO 680, and the oxazine derivative MR 121 are strongly quenched almost exclusively by tryptophan due to the formation of weak or nonfluorescent ground-state complexes with association constants, K-ass., ranging from 96 to 206 M-1. Rhodamine, fluorescein, and bora-diaza-indacene derivatives that absorb at shorter wavelengths are also quenched substantially by tyrosine residues. The quenching of carbocyanine dyes, such as Cy5, and Alexa 647 by amino acids can be almost neglected. While quenching of ATTO 655, ATTO 680, and the oxazine derivative MR121 by tryptophan is dominated by static quenching, dynamic quenching is more efficient for the two bora-diaza-indacene dyes Bodipy-FL and Bodipy630/650. Labeling of the dyes to tryptophan, tryptophan-containing peptides, and proteins (streptavidin) demonstrates that knowledge of these fluorescence quenching processes is crucial for the development of fluorescence-based diagnostic assays. Changes in the fluorescence quantum yield of dye-labeled peptides and proteins might be used advantageously for the quantification of proteases and specific binding partners.
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页码:1133 / 1139
页数:7
相关论文
共 33 条
  • [1] Charge transfer on the nanoscale: Current status
    Adams, DM
    Brus, L
    Chidsey, CED
    Creager, S
    Creutz, C
    Kagan, CR
    Kamat, PV
    Lieberman, M
    Lindsay, S
    Marcus, RA
    Metzger, RM
    Michel-Beyerle, ME
    Miller, JR
    Newton, MD
    Rolison, DR
    Sankey, O
    Schanze, KS
    Yardley, J
    Zhu, XY
    [J]. JOURNAL OF PHYSICAL CHEMISTRY B, 2003, 107 (28) : 6668 - 6697
  • [2] DETECTION OF NUCLEIC-ACID HYBRIDIZATION BY NONRADIATIVE FLUORESCENCE RESONANCE ENERGY-TRANSFER
    CARDULLO, RA
    AGRAWAL, S
    FLORES, C
    ZAMECNIK, PC
    WOLF, DE
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (23) : 8790 - 8794
  • [3] Biotin and digoxigenin as labels for light and electron microscopy in situ hybridization probes: Where do we stand?
    Chevalier, J
    Yi, J
    Michel, O
    Tang, XM
    [J]. JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1997, 45 (04) : 481 - 491
  • [4] ELECTROCHEMICAL PROPERTIES OF TYROSINE PHENOXY AND TRYPTOPHAN INDOLYL RADICALS IN PEPTIDES AND AMINO-ACID-ANALOGS
    DEFELIPPIS, MR
    MURTHY, CP
    BROITMAN, F
    WEINRAUB, D
    FARAGGI, M
    KLAPPER, MH
    [J]. JOURNAL OF PHYSICAL CHEMISTRY, 1991, 95 (08) : 3416 - 3419
  • [5] Conformational transitions monitored for single molecules in solution
    Edman, L
    Mets, U
    Rigler, R
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (13) : 6710 - 6715
  • [6] Monitoring conformational dynamics of a single molecule by selective fluorescence spectroscopy
    Eggeling, C
    Fries, JR
    Brand, L
    Günther, R
    Seidel, CAM
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (04) : 1556 - 1561
  • [7] IMAGING OF ENDOSOME FUSION IN BHK FIBROBLASTS BASED ON A NOVEL FLUOROMETRIC AVIDIN-BIOTIN BINDING ASSAY
    EMANS, N
    BIWERSI, J
    VERKMAN, AS
    [J]. BIOPHYSICAL JOURNAL, 1995, 69 (02) : 716 - 728
  • [8] FORSTER T, 1949, Z NATURFORSCH A, V4, P321
  • [9] Quantitative fluorescence resonance energy transfer measurements using fluorescence microscopy
    Gordon, GW
    Berry, G
    Liang, XH
    Levine, B
    Herman, B
    [J]. BIOPHYSICAL JOURNAL, 1998, 74 (05) : 2702 - 2713
  • [10] Anomalous fluorescence enhancement of Cy3 and Cy3.5 versus anomalous fluorescence loss of Cy5 and Cy7 upon covalent linking to IgG and noncovalent binding to avidin
    Gruber, HJ
    Hahn, CD
    Kada, G
    Riener, CK
    Harms, GS
    Ahrer, W
    Dax, TG
    Knaus, HG
    [J]. BIOCONJUGATE CHEMISTRY, 2000, 11 (05) : 696 - 704