cat-ELCCA: A Robust Method To Monitor the Fatty Acid Acyltransferase Activity of Ghrelin O-Acyltransferase (GOAT)

Authors

  • Dr. Amanda L. Garner,

    1. Departments of Chemistry and Immunology, The Skaggs Institute for Chemical Biology and The Worm Institute for Research and Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-2595
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  • Prof. Kim D. Janda

    Corresponding author
    1. Departments of Chemistry and Immunology, The Skaggs Institute for Chemical Biology and The Worm Institute for Research and Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-2595
    • Departments of Chemistry and Immunology, The Skaggs Institute for Chemical Biology and The Worm Institute for Research and Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-2595
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  • This work was supported by the Skaggs Institute for Chemical Biology (K.D.J.) and by the NIH (postdoctoral fellowship F32-DK083179 to A.L.G.). We are grateful to Damian Ekiert for assistance with baculovirus expression, Prof. M. G. Finn for helpful discussions, and Vu Hong for donation of THPTA.

Abstract

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Assays armed with catalytic signal amplification have arisen as superior systems for ultrasensitive detection of analytes. A conceptually new enzyme assay called cat-ELCCA (catalytic assay using enzyme-linked click-chemistry) is described, in which an enzyme-linked azide is utilized to arm the assay with catalytic fluorescence signal amplification. By using this assay technology, the first high-throughput screen for recently disclosed ghrelin O-acyltransferase (GOAT) was developed.

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