Skip to Content
Merck
  • Type I phosphatidylinositol 4-phosphate 5-kinase homo- and heterodimerization determines its membrane localization and activity.

Type I phosphatidylinositol 4-phosphate 5-kinase homo- and heterodimerization determines its membrane localization and activity.

FASEB journal : official publication of the Federation of American Societies for Experimental Biology (2015-02-26)
Rosa Ana Lacalle, Juan C de Karam, Laura Martínez-Muñoz, Ibai Artetxe, Rosa M Peregil, Jesús Sot, Ana M Rojas, Félix M Goñi, Mario Mellado, Santos Mañes
ABSTRACT

Type I phosphatidylinositol 4-phosphate 5-kinases (PIP5KIs; α, β, and γ) are a family of isoenzymes that produce phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] using phosphatidylinositol 4-phosphate as substrate. Their structural homology with the class II lipid kinases [type II phosphatidylinositol 5-phosphate 4-kinase (PIP4KII)] suggests that PIP5KI dimerizes, although this has not been formally demonstrated. Neither the hypothetical structural dimerization determinants nor the functional consequences of dimerization have been studied. Here, we used Förster resonance energy transfer, coprecipitation, and ELISA to show that PIP5KIβ forms homo- and heterodimers with PIP5KIγ_i2 in vitro and in live human cells. Dimerization appears to be a general phenomenon for PIP5KI isoenzymes because PIP5KIβ/PIP5KIα heterodimers were also detected by mass spectrometry. Dimerization was independent of actin cytoskeleton remodeling and was also observed using purified proteins. Mutagenesis studies of PIP5KIβ located the dimerization motif at the N terminus, in a region homologous to that implicated in PIP4KII dimerization. PIP5KIβ mutants whose dimerization was impaired showed a severe decrease in PI(4,5)P2 production and plasma membrane delocalization, although their association to lipid monolayers was unaltered. Our results identify dimerization as an integral feature of PIP5K proteins and a central determinant of their enzyme activity.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Tris(tert-butoxy)silanol, 99.999%
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Guanosine, ≥97.0% (HPLC)
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
8-Octanoyloxypyrene-1,3,6-trisulfonic acid trisodium salt, suitable for fluorescence, ≥90% (HPCE)
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Dimethyl sulfoxide, BioUltra, for molecular biology, ≥99.5% (GC)
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Dimethyl sulfoxide, anhydrous, ≥99.9%
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
N-Formyl-Met-Leu-Phe, BioXtra, ≥99.0% (TLC)
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
L-Glutathione reduced, BioXtra, ≥98.0%
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
L-Glutathione reduced, suitable for cell culture, BioReagent, ≥98.0%, powder
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Guanosine, BioReagent, suitable for cell culture
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Guanosine, ≥98%
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
N-Formyl-Met-Leu-Phe, ≥97% (HPLC)
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Dimethyl sulfoxide, PCR Reagent
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
L-Glutathione reduced, ≥98.0%
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Dimethyl sulfoxide, meets EP testing specifications, meets USP testing specifications
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Dimethyl sulfoxide, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Dimethyl sulfoxide, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Dimethyl sulfoxide, for molecular biology
Pricing and Inventory is not available. Please check the Merck website for more information.
Sigma-Aldrich
Dimethyl sulfoxide, ≥99.5% (GC), suitable for plant cell culture
Pricing and Inventory is not available. Please check the Merck website for more information.