Fluorescent detection of proteins in polyacrylamide gels

Details Fluorescent detection of proteins in polyacrylamide gels Fluorescent detection of proteins in polyacrylamide gels

2003-01-27

2009-08-04

Barton, Jeffrey (Department: 1795)

Chemistry: electrical and wave energy

Processes and products

Electrophoresis or electro-osmosis processes and electrolyte...

C204S469000, C204S470000

Reexamination Certificate

active

07569130

ABSTRACT:
The mechanism of the UV light-induced reaction between the indole moiety of tryptophan and chloroform, and the structure of the modified tryptophan and polypeptides including such modified tryptophan residues. The excited indole moiety, which is formed upon UV light irradiation, emits a solvated electron which initiates a series of events that yield fluorescent derivatives that have CHO group covalently bound to the indole moiety. These derivatives are herein referred to as formyltryptophan, and are relatively stable. Similar reactions are observed when 5-hydroxytryptophan, 5-fluorotryptophan, or N-methylindolacetate are used in place of tryptophan, or when other haloalkanes, such as trichloracetic acid, trichlorethanol, trichlorethane, bromoform, and iodoactetate are used in place of chloroform. The derivatives can be used in a variety of applications in fluorescence spectroscopy, and for nuclear magnetic resonance, X-ray crystallography, infra-red spectroscopy, circular dicroism and mass spectroscopy. Additionally, the UV light-induced reaction between the indole moiety of tryptophan and haloalkanes can be used to prepare derivatives of tryptophan for chemical cross-linking studies of proteins and peptides.

REFERENCES:
patent: 4119521 (1978-10-01), Chirikjian
patent: 5865975 (1999-02-01), Bishop
J.-P. Privat et al, “Photochemical Modifications of the Tryptophan Residues of Wheat-Germ Agglutinin in the Presence of Trichloroethanol” Eur. J. Biochem. 84, 79-85. (1978).
E. Mehl, “Separation and Characterization of Myelin Proteins”, Advances in Experimental Medicine and Biology. 32, 157-170. (1972).
“SYPRO Orange and Red protein gel stains” brochure from Amersham Biosciences. 4 pages, published in 2000.
Edwards, et al. “Visualization of Proteins in Acrylamide Gels Using Ultraviolet Illumination” Analytical Biochemistry 301, 91-96 (2002) Article.
Vorpel, et al. “Chloroform-Dependent Photoproducts of Tryptophan”, Biophysics, vol. 37, No. 5, pp. 743-745, 1992—Article.
Pinchuk. et al. Spctral Characteristics and Mechanisms of Forming “Choloroform-Dependent Tryptophan Photoproducts”, Institute of Photobiology, vol. 59. Nos. 3-4 pp. 312-317, 1993—Article.
Wong, et al., “Heating Greatly Speeds Coomassie Blue Staining and Destaining”, BioTechniques 28:426-432 (Mar. 2000)—Article.
Patton, Wayne F. “A thousand points of light: The Application of fluorescence detection technologies to two-dimensional gel electrophoresis and proteomics”, Electrophoresis 2000, 21, 1123-1144—Article.
Berggren, et al. “Background-free, high sensitivity staining of proteins in one-and two-dimensional sodium dodecyl sulfate-polyacrylamide gels, etc.” Electrophoresis 2000, 21, 2509-2521—Article.
Yamamoto H. et al.: “Ultraviolet Imaging Densitometry of Unstained Gels for Two-Dimensional Electrophoresis”, Anatytical Biochemistry. United States Nov. 15, 1990, vol. 191, No. 1, pp. 58-64.
Roegener Jan, et al. “Ultrasensitive Detection of Unstained Proteins in Acrylamide Gels by Native UV Fluorescence”, Analytical Chem., U.S. Jan. 1, 2003—vol. 75, No. 1 pp. 157-159.
Edwards, Robert A. et al. “The Light-Induced Reactions of Tryptophan With Halocompounds” Photochemistry and Photobiology. U.S. Apr. 2002, vol. 75, No. 4, pp. 362-368.
Leibowitz M J et al. “Visualization and Elution of Unstained Proteins from Polyacrylamide Gels” Analytical Biochem. U.S. Feb. 1984, vol. 137, No. 1, pp. 161-163.
Koutny, Lance B. et al: “On-line detection of proteins in gel electrophoresis by ultraviolet absorption and by native fluorescence utilizing a charge-coupled device imaging system”—Anal. Chem. vo. 65, No. 2, 1993, pp. 183-187.

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