Dimerization and phosphorylation of thyrotropin-releasing hormone receptors are modulated by agonist stimulation

Changcheng Zhu, Laurie B. Cook, Patricia M. Hinkle

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Dimerization and phosphorylation of thyrotropin-releasing hormone (TRH) receptors was characterized using HEK293 and pituitary GHFT cells expressing epitope-tagged receptors. TRH receptors tagged with FLAG and hemagglutinin epitopes were co-precipitated only if they were co-expressed, and 10-30% of receptors were isolated as hemagglutinin/FLAG-receptor dimers under basal conditions. The abundance of receptor dimers was increased when cells had been stimulated by TRH, indicating that TRH either stabilizes pre-existing dimers or increases dimer formation. TRH increased receptor dimerization and phosphorylation within 1 min in a dose-dependent manner. TRH increased phosphorylation of both receptor monomers and dimers, documented by incorporation of 32P and an upshift in receptor mobility reversed by phosphatase treatment. The ability of TRH to increase receptor phosphorylation and dimerization did not depend on signal transduction, because it was not inhibited by the phospholipase C inhibitor U73122. Receptor phosphorylation required an agonist but was not blocked by the casein kinase II inhibitor apigenin, the protein kinase C inhibitor GF109203X, or expression of a dominant negative form of G protein-coupled receptor kinase 2. TRH receptors lacking most of the cytoplasmic carboxyl terminus formed dimers constitutively but failed to undergo agonist-induced dimerization and phosphorylation. TRH also increased phosphorylation and dimerization of TRH receptors expressed in GHFT pre-lactotroph cells.

Original languageEnglish (US)
Pages (from-to)28228-28237
Number of pages10
JournalJournal of Biological Chemistry
Volume277
Issue number31
DOIs
StatePublished - Aug 2 2002
Externally publishedYes

Fingerprint

Thyrotropin Releasing Hormone Receptors
Phosphorylation
Dimerization
Thyrotropin-Releasing Hormone
Dimers
Hormones
Hemagglutinins
Epitopes
G-Protein-Coupled Receptor Kinase 2
Lactotrophs
Apigenin
Casein Kinase II
Signal transduction
Protein C Inhibitor
Type C Phospholipases
Protein Kinase Inhibitors
Phosphoric Monoester Hydrolases
Protein Kinase C
Signal Transduction
Monomers

ASJC Scopus subject areas

  • Biochemistry

Cite this

Dimerization and phosphorylation of thyrotropin-releasing hormone receptors are modulated by agonist stimulation. / Zhu, Changcheng; Cook, Laurie B.; Hinkle, Patricia M.

In: Journal of Biological Chemistry, Vol. 277, No. 31, 02.08.2002, p. 28228-28237.

Research output: Contribution to journalArticle

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