title: Regulation of phospholipase C-beta 3 activity by Na+/H+ exchanger regulatory factor 2.
abstract: Among the phospholipase C that catalyzes the hydrolysis of phosphatidylinositol 4,5-bisphosphate, four mammalian phospholipase C-beta (PLC-beta) isotypes (isotypes 1-4) are activated through G protein-coupled receptors (GPCRs). Although the regulation of the PLC-betas by GPCRs and heterotrimeric G proteins has been extensively studied, little is known about the molecular determinants that regulate their activity. The PLC-beta isozymes carry a putative PSD-95/Dlg/ZO-1 (PDZ) binding motif (X(S/T)X(V/L)COOH) at their carboxyl terminus, which is implicated in specific interactions with anchor proteins. Using the yeast two-hybrid system, we identified Na(+)/H(+) exchanger regulatory factor 2 (NHERF2) as a protein that interacted with a C-terminal heptapeptide of PLC-beta3. Immunoprecipitation studies revealed that NHERF2 interacts specifically with PLC-beta3, but not with other PLC-beta isotypes. Furthermore, PLC-beta3 interacted with NHERF2 rather than with other PDZ-containing proteins. This interaction required the COOH-terminal NTQL sequence of PLC-beta3 and the second PDZ domain of NHERF2. Interestingly, NHERF2 potentiated the PLC-beta activation by carbachol in COS7 and HeLa cells, while mutant NHERF2, lacking the second PDZ domain, had no such effect. Taken together, the data suggest that NHERF2 may act as a modulator underlying the process of PLC-beta3-mediated signaling.
authors.affiliation: Department of Life Science, National Creative Research Initiative Center for Calcium and Learning, Division of Molecular and Life Science and School of Environmental Engineering, Pohang University of Science and Technology, Pohang 790-784, South Korea.
authors.forename: J I
authors.initials: JI
authors.lastname: Hwang
authors.forename: K
authors.initials: K
authors.lastname: Heo
authors.forename: K J
authors.initials: KJ
authors.lastname: Shin
authors.forename: E
authors.initials: E
authors.lastname: Kim
authors.forename: C
authors.initials: C
authors.lastname: Yun
authors.forename: S H
authors.initials: SH
authors.lastname: Ryu
authors.forename: H S
authors.initials: HS
authors.lastname: Shin
authors.forename: P G
authors.initials: PG
authors.lastname: Suh
mesh_terms: D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism*; D007930:Leucine / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism; D054799:Phospholipase C beta; D010750:Phosphoproteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D015398:Signal Transduction; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*; D013379:Substrate Specificity; D013912:Threonine / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D010738:Type C Phospholipases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D007527:Isoenzymes; D010716:Phosphatidylinositols; D010750:Phosphoproteins; D017923:Sodium-Hydrogen Exchangers; C093376:sodium-hydrogen exchanger regulatory factor; D013912:Threonine; D010738:Type C Phospholipases; C518039:PLCB3 protein, human; D054799:Phospholipase C beta; D007930:Leucine