Modified fluorescent proteins for detecting protease activity

Details Modified fluorescent proteins for detecting protease activity Modified fluorescent proteins for detecting protease activity

2006-05-23

2006-05-23

Saidha, Tekchand (Department: 1652)

Chemistry: natural resins or derivatives; peptides or proteins;

Proteins, i.e., more than 100 amino acid residues

C435S183000, C435S212000, C435S219000, C435S252300, C435S320100

Reexamination Certificate

active

07049400

ABSTRACT:
The present invention concerns fluorescent proteins modified such that said modified fluorescent protein incorporates a cleavage site for a protease, cleavage of said modified fluorescent protein at said cleavage site by said protease causing the alteration of at least one of the emission and excitation spectra of said modified fluorescent protein. In particular, the invention is concerned with using these modified fluorescent proteins as probes for detecting protease activity in living cells during the programmed cell death process (apoptosis).Also provided are nucleic acid sequence encoding same, recombinant DNA constructs expressing same, cells transformed or transfected with same, methods for detecting protease activity, and methods of detecting agents which affect protease activity, and kits for same.

REFERENCES:
patent: 5491084 (1996-02-01), Chalfie et al.
patent: 5625048 (1997-04-01), Tsien et al.
patent: 5777079 (1998-07-01), Tsien et al.
patent: 5804387 (1998-09-01), Cormack et al.
patent: 5981200 (1999-11-01), Tsien et al.
Branden et al.. Introduction to Protein Structure. New York and London: Garland Publishing, Inc. 1991, p. 271.
Xu et al. Detection of programmed cell death using fluorescence energy transfer. Nucleic Acids Res. Apr. 15, 1998;26(8):2034-5.
Cody et al. Biochemistry. Feb. 9, 1993;32(5):1212-8 (ABSTRACT).
Thornberry et al, “A Combinatorial Approach Defines Specificites of Membes of the Caspase Family and Cranzyme B”,The Journal of Biological Chemistry, vol. 272, No. 29, Issue of Jul. 18, 1997, pp. 17907-17911, USA.
“ApoAlert™ CPP32 Protease Assay Kits”, CLONTECHniques, Jan., 1997, pp. 4-6, USA.
Geoffrey S. Braid et al, “Circular permutation and receptor insertion within green fluorescent proteins,”Proc. Natl. Acad. Sci. USA,vol. 96, pp. 11241-11246, Sep., 1999, US.
Mats Ormo et al, “Crystal Structure of theQequorea victoriaGreen Fluorescent Protein”,Science,vol. 273, pp. 1392-1395, Sep. 6, 1996, US.
Fan Yang et al, “The molecular structure of green fluorescent protein”,Nature Biotechnology,vol. 14, pp. 1246-1251, Oct. 14, 1996, US.
Rebekka M. Wachter et al, “Crystal Structure and Photodynamic Behavior of the Blue Emission Variant Y66H/Y145F of Green Fluorescent Protein”,Biochemistry,vol. 36, pp. 9759-9765, 1997, American Chemical Society, US.
Daniel Yarbrough et al, “Refined crystal structure of DsRed, a red fluorescent protein from coral, at 2.0-A resolution”,Proc Natl Acad Sci USA,vol. 98, pp. 462-467, Jan. 16, 2001, US.
Abstract of Article—Engineering green fluorescent protein for improved brightness, longer wavelength and fluorescence resonance energy transfer,R. Heim and T. Y. Tsien, Curr. Biol., Feb. 1996, 1 page.
Article—Detection of programmed cell death using fluorescence energy transfer,Xiang Xu, Amy L. V. Gerard, Betty C. B. Huang, David C. Anderson, Donald G. Payan, and Ying Luo, Nucleic Acids Research, vol. 26, No. 8, 1998, pp. 2034-2035.
Article—Dual Labeling Using ECFP&EYFP in Standard Fluorescence Microscopy,Brigitte Angres & Gisele Green, CLONTECH, Apr. 1999, 2 pages.
Article—Understanding, improving and using green fluorescent proteins,Andrew B. Cubitt, Roger Heim, Stephen R. Adams, Aileen E. Boyd, Larry A. Gross, and Roger Y. Tsien, Techniques, TIBS 20, Nov. 1995, pp. 448-455.
Product Information on ApoAlert™ Caspase Assay Kits from CLONTECH, 1 page.
Product Information on Living Colors™ pEBFP-N1 & -C1 Vectors from CLONTECH, 2 pages.
Query Result Browser on Green Fluorescent Protein from Protein Data Bank, 4 pages, www.rcsb.org.
Summary Information—Blue Variant Of Green Fluorescent Protein by R. M. Wachter, and S. J. Remington from Protein Data Bank, Deposition Date: Apr. 9, 1997, www.rcsb.org.
Summary Information—Green Fluorescent Protein (Gfp) FromAequorea victoria,Gln 80 Replaced With Arg by K. Brejc and T. Sixma from Protein Data Bank, Deposition Date: Jan. 8, 1997, www.rcsb.org.
Summary Information—Green Fluorescent Protein FromAequorea victoriaby M. Ormo and S. J. Remington from Protein Data Bank, Deposition Date: Aug. 1, 1996, www.rcsb.org.
Summary Information—Green Fluorescent Protein FromAequorea victoria,Mutant by G. Palm, Z. Zdanov, and A. Wlodawer from Protein Data Bank, Deposition Date: Mar. 31, 1997, www.rcsb.org.
Summary Information—Green Fluorescent Protein Mutant F99S, M153T and V163A by R. Battistutta, A. Negro, and G. Zanotti from Protein Data Bank, Deposition Date: Feb. 9, 1999, www.rcsb.org.
Summary Information—Green Fluorescent Protein S65T At pH 4.6 by M. A. Elsliger, R. M. Wachter, K. Kallio, G. T. Hanson, and S. J. Remington from Protein Data Bank, Deposition Date: Aug. 21, 1999, www.rcsb.org.
Summary Information—Structure Of Yellow-Emission Variant Of Gfp by R. M. Wachter, M.-A. Elsliger, K. Kallio, G. T. Hanson, and S. J. Remington from Protein Data Bank, Deposition Date: Aug. 28, 1998, www.rcsb.org.

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