Measurement of changes in fluorescence resonance energy transfer between gonadotropin-releasing hormone receptors in response to agonists

Anda Cornea, P. Michael Conn

    Research output: Contribution to journalArticle

    15 Citations (Scopus)

    Abstract

    Oligomerization of membrane-bound G-protein-coupled receptors has recently emerged as an important step in cellular signaling. Fluorescence resonance energy transfer (FRET) has undergone a revival as the method of choice for demonstrating in vivo protein-protein interactions and receptor dimerization. We have used chimeras of gonadotropin-releasing harmone (GnRH) receptors and various fluorescent proteins to investigate receptor dimerization in relation to receptor activation. Two pairs of FRET-compatible fluorescent proteins were used: sapphire with topaz, and enhanced green fluorescent protein (eGFP) with dsRed. Changes in the ratio between acceptor and donor fluorescence were measured after addition of buserelin, a GnRH agonist, and antide, a GnRH antagonist. For both pairs of fluorescent proteins, an increase in the ratio of acceptor to donor intensities was observed immediately after addition of buserelin as would be predicted if FRET occurred due to the microaggregation of receptors conjugated with different fluorescent proteins. No change in FRET was observed in time for cells in medium or after addition of antide. The increase in FRET signal was not uniform throughout a cell.

    Original languageEnglish (US)
    Pages (from-to)333-339
    Number of pages7
    JournalMethods
    Volume27
    Issue number4
    DOIs
    StatePublished - 2002

    Fingerprint

    LHRH Receptors
    Fluorescence Resonance Energy Transfer
    Gonadotropins
    Buserelin
    Proteins
    Dimerization
    Topaz
    Cell signaling
    Oligomerization
    Aluminum Oxide
    G-Protein-Coupled Receptors
    Fluorescence
    Chemical activation
    Membranes

    ASJC Scopus subject areas

    • Molecular Biology

    Cite this

    Measurement of changes in fluorescence resonance energy transfer between gonadotropin-releasing hormone receptors in response to agonists. / Cornea, Anda; Michael Conn, P.

    In: Methods, Vol. 27, No. 4, 2002, p. 333-339.

    Research output: Contribution to journalArticle

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