Cover Picture: A TR-FRET-Based Functional Assay for Screening Activators of CARM1 (ChemBioChem 7/2013)

Authors

  • Hao Zeng,

    1. Graduate Program in Cellular and Molecular Biology, McArdle Laboratory for Cancer Research and Carbone Comprehensive Cancer Center, University of Wisconsin, 1400 University Ave, Madison, WI 53706 (USA)
    Search for more papers by this author
  • Dr. Jiacai Wu,

    1. Graduate Program in Cellular and Molecular Biology, McArdle Laboratory for Cancer Research and Carbone Comprehensive Cancer Center, University of Wisconsin, 1400 University Ave, Madison, WI 53706 (USA)
    2. Current address: The Center for Science Research, Institute of Biomedical Sciences, Guilin Medical University, Guilin, Guangxi, 541004 (P.R. China)
    Search for more papers by this author
  • Prof. Dr. Mark T. Bedford,

    1. Department of Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, TX 78957 (USA)
    Search for more papers by this author
  • Prof. Dr. Gianluca Sbardella,

    1. Dipartimento di Farmacia, Epigenetic Med Chem Lab, Università degli Studi di Salerno, Via Ponte Don Melillo, 84084 Fisciano (SA) (Italy)
    Search for more papers by this author
  • Prof. Dr. F. Michael Hoffmann,

    1. Graduate Program in Cellular and Molecular Biology, McArdle Laboratory for Cancer Research and Carbone Comprehensive Cancer Center, University of Wisconsin, 1400 University Ave, Madison, WI 53706 (USA)
    Search for more papers by this author
  • Dr. Kun Bi,

    1. Life Technologies Corporation, Primary and Stem Cell Systems, Madison, WI 53719 (USA)
    Search for more papers by this author
  • Prof. Dr. Wei Xu

    Corresponding author
    1. Graduate Program in Cellular and Molecular Biology, McArdle Laboratory for Cancer Research and Carbone Comprehensive Cancer Center, University of Wisconsin, 1400 University Ave, Madison, WI 53706 (USA)
    • Graduate Program in Cellular and Molecular Biology, McArdle Laboratory for Cancer Research and Carbone Comprehensive Cancer Center, University of Wisconsin, 1400 University Ave, Madison, WI 53706 (USA)

    Search for more papers by this author

Abstract

original image

The cover picture shows a TR-FRET-based functional assay for screening small-molecule modulators of co-activator-associated arginine methyltransferase 1 (CARM1), based on its methylation of PABP1. The formation of the ternary complex (Me-GFP-PABP1, Me-PABP1 Ab, and Tb-2nd Ab) enables time-resolved detection of the long-lifetime terbium (Tb)-to-GFP FRET signal. Light at 340 nm is used to excite Tb, which emits at 495 nm. The proximity of the donor Tb to the acceptor GFP results in energy transfer from Tb to GFP, and this induces the specific FRET signal at 520 nm. On p. 827 ff., W. Xu et al. describe the development and optimization of the homogenous Me-GFP-PABP1-based TR-FRET assay for monitoring the expression and enzymatic activity of CARM1 in MCF7 breast cancer cells. The assay has been optimized for high-throughput screening to identify CARM1 allosteric activators. More importantly, this methodology has the potential to be extended to other protein arginine methyltransferases through the development of appropriate substrate–GFP fusion proteins and methylation-site-specific antibodies.

Cartoon 1.

Ancillary