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The pleckstrin homology domain of phosphoinositide-specific phospholipase Cdelta4 is not a critical determinant of the membrane localization of the enzyme.
J Biol Chem. 2004 Jun 04; 279(23):24362-71.JB

Abstract

The inositol lipid and phosphate binding properties and the cellular localization of phospholipase Cdelta(4) (PLCdelta(4)) and its isolated pleckstrin homology (PH) domain were analyzed in comparison with the similar features of the PLCdelta(1) protein. The isolated PH domains of both proteins showed plasma membrane localization when expressed in the form of a green fluorescent protein fusion construct in various cells, although a significantly lower proportion of the PLCdelta(4) PH domain was membrane-bound than in the case of PLCdelta(1)PH-GFP. Both PH domains selectively recognized phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)), but a lower binding of PLCdelta(4)PH to lipid vesicles containing PI(4,5)P(2) was observed. Also, higher concentrations of inositol 1,4,5-trisphosphate (Ins(1,4,5)P(3)) were required to displace the PLCdelta(4)PH from the lipid vesicles, and a lower Ins(1,4,5)P(3) affinity of PLCdelta(4)PH was found in direct Ins(1,4,5)P(3) binding assays. In sharp contrast to the localization of its PH domain, the full-length PLCdelta(4) protein localized primarily to intracellular membranes mostly to the endoplasmic reticulum (ER). This ER localization was in striking contrast to the well documented PH domain-dependent plasma membrane localization of PLCdelta(1). A truncated PLCdelta(4) protein lacking the entire PH domain still showed the same ER localization as the full-length protein, indicating that the PH domain is not a critical determinant of the localization of this protein. Most important, the full-length PLCdelta(4) enzyme still showed binding to PI(4,5)P(2)-containing micelles, but Ins(1,4,5)P(3) was significantly less potent in displacing the enzyme from the lipid than with the PLCdelta(1) protein. These data suggest that although structurally related, PLCdelta(1) and PLCdelta(4) are probably differentially regulated in distinct cellular compartments by PI(4,5)P(2) and that the PH domain of PLCdelta(4) does not act as a localization signal.

Authors+Show Affiliations

Laboratory of Cell Signaling, NHLI, National Institutes of Health, Bethesda, Maryland 20892, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15037625

Citation

Lee, Sang Bong, et al. "The Pleckstrin Homology Domain of Phosphoinositide-specific Phospholipase Cdelta4 Is Not a Critical Determinant of the Membrane Localization of the Enzyme." The Journal of Biological Chemistry, vol. 279, no. 23, 2004, pp. 24362-71.
Lee SB, Várnai P, Balla A, et al. The pleckstrin homology domain of phosphoinositide-specific phospholipase Cdelta4 is not a critical determinant of the membrane localization of the enzyme. J Biol Chem. 2004;279(23):24362-71.
Lee, S. B., Várnai, P., Balla, A., Jalink, K., Rhee, S. G., & Balla, T. (2004). The pleckstrin homology domain of phosphoinositide-specific phospholipase Cdelta4 is not a critical determinant of the membrane localization of the enzyme. The Journal of Biological Chemistry, 279(23), 24362-71.
Lee SB, et al. The Pleckstrin Homology Domain of Phosphoinositide-specific Phospholipase Cdelta4 Is Not a Critical Determinant of the Membrane Localization of the Enzyme. J Biol Chem. 2004 Jun 4;279(23):24362-71. PubMed PMID: 15037625.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The pleckstrin homology domain of phosphoinositide-specific phospholipase Cdelta4 is not a critical determinant of the membrane localization of the enzyme. AU - Lee,Sang Bong, AU - Várnai,Péter, AU - Balla,Andras, AU - Jalink,Kees, AU - Rhee,Sue-Goo, AU - Balla,Tamas, Y1 - 2004/03/22/ PY - 2004/3/24/pubmed PY - 2004/7/9/medline PY - 2004/3/24/entrez SP - 24362 EP - 71 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 279 IS - 23 N2 - The inositol lipid and phosphate binding properties and the cellular localization of phospholipase Cdelta(4) (PLCdelta(4)) and its isolated pleckstrin homology (PH) domain were analyzed in comparison with the similar features of the PLCdelta(1) protein. The isolated PH domains of both proteins showed plasma membrane localization when expressed in the form of a green fluorescent protein fusion construct in various cells, although a significantly lower proportion of the PLCdelta(4) PH domain was membrane-bound than in the case of PLCdelta(1)PH-GFP. Both PH domains selectively recognized phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)), but a lower binding of PLCdelta(4)PH to lipid vesicles containing PI(4,5)P(2) was observed. Also, higher concentrations of inositol 1,4,5-trisphosphate (Ins(1,4,5)P(3)) were required to displace the PLCdelta(4)PH from the lipid vesicles, and a lower Ins(1,4,5)P(3) affinity of PLCdelta(4)PH was found in direct Ins(1,4,5)P(3) binding assays. In sharp contrast to the localization of its PH domain, the full-length PLCdelta(4) protein localized primarily to intracellular membranes mostly to the endoplasmic reticulum (ER). This ER localization was in striking contrast to the well documented PH domain-dependent plasma membrane localization of PLCdelta(1). A truncated PLCdelta(4) protein lacking the entire PH domain still showed the same ER localization as the full-length protein, indicating that the PH domain is not a critical determinant of the localization of this protein. Most important, the full-length PLCdelta(4) enzyme still showed binding to PI(4,5)P(2)-containing micelles, but Ins(1,4,5)P(3) was significantly less potent in displacing the enzyme from the lipid than with the PLCdelta(1) protein. These data suggest that although structurally related, PLCdelta(1) and PLCdelta(4) are probably differentially regulated in distinct cellular compartments by PI(4,5)P(2) and that the PH domain of PLCdelta(4) does not act as a localization signal. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/15037625/The_pleckstrin_homology_domain_of_phosphoinositide_specific_phospholipase_Cdelta4_is_not_a_critical_determinant_of_the_membrane_localization_of_the_enzyme_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=15037625 DB - PRIME DP - Unbound Medicine ER -