Phospholipase C-β3 and -β1 form homodimers, but not heterodimers, through catalytic and carboxyl-terminal domains

Yong Zhang, Walter K. Vogel, Jennifer S. McCullar, Jeffrey A. Greenwood, Theresa M. Filtz

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Phospholipase C-β (PLC-β) isoenzymes are key effectors in G protein-coupled signaling pathways. Prior research suggests that some isoforms of PLC-β may exist and function as dimers. Using coimmunoprecipitation assays of differentially tagged PLC-β constructs and size-exclusion chromatography of native PLC-β, we observed homodimerization of PLC-β3 and PLC-β1 isoenzymes but failed to detect heterodimerization of these isoenzymes. Size-exclusion chromatography data suggest that PLC-β3 and PLC-β1 form higher affinity homodimers than PLC-β2. Evidence supportive of limited PLC-β monomer-homodimer equilibrium appears at ≤100 nM. Further assessment of homodimerization status by coimmunoprecipitation assays with differentially tagged PLC-β3 fragments demonstrated that at least two subdomains of PLC-β3 are involved in dimer formation, one in the catalytic X and Y domains and the other in the G protein-regulated carboxyl-terminal domain. In addition, we provide evidence consistent with the existence of PLC-β homodimers in a whole-cell context, using fluorescent protein-tagged constructs and microscopic fluorescence resonance energy transfer assays.

Original languageEnglish (US)
Pages (from-to)860-868
Number of pages9
JournalMolecular Pharmacology
Volume70
Issue number3
DOIs
StatePublished - Aug 28 2006
Externally publishedYes

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Type C Phospholipases
Isoenzymes
GTP-Binding Proteins
Gel Chromatography
Fluorescence Resonance Energy Transfer
Protein Isoforms

ASJC Scopus subject areas

  • Pharmacology

Cite this

Phospholipase C-β3 and -β1 form homodimers, but not heterodimers, through catalytic and carboxyl-terminal domains. / Zhang, Yong; Vogel, Walter K.; McCullar, Jennifer S.; Greenwood, Jeffrey A.; Filtz, Theresa M.

In: Molecular Pharmacology, Vol. 70, No. 3, 28.08.2006, p. 860-868.

Research output: Contribution to journalArticle

Zhang, Yong ; Vogel, Walter K. ; McCullar, Jennifer S. ; Greenwood, Jeffrey A. ; Filtz, Theresa M. / Phospholipase C-β3 and -β1 form homodimers, but not heterodimers, through catalytic and carboxyl-terminal domains. In: Molecular Pharmacology. 2006 ; Vol. 70, No. 3. pp. 860-868.
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