ORP2 interacts with phosphoinositides and controls the subcellular distribution of cholesterol

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http://hdl.handle.net/10138/299791

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Koponen , A , Arora , A , Takahashi , K , Kentala , H , Kivelä , A M , Jääskeläinen , E , Peränen , J , Somerharju , P , Ikonen , E , Viitala , T & Olkkonen , V M 2019 , ' ORP2 interacts with phosphoinositides and controls the subcellular distribution of cholesterol ' , Biochimie , vol. 158 , pp. 90-101 . https://doi.org/10.1016/j.biochi.2018.12.013

Title: ORP2 interacts with phosphoinositides and controls the subcellular distribution of cholesterol
Author: Koponen, Annika; Arora, Amita; Takahashi, Kohta; Kentala, Henriikka; Kivelä, Annukka M.; Jääskeläinen, Eeva; Peränen, Johan; Somerharju, Pentti; Ikonen, Elina; Viitala, Tapani; Olkkonen, Vesa M.
Contributor: University of Helsinki, Minerva Foundation Institute for Medical Research
University of Helsinki, Department of Anatomy
University of Helsinki, Department of Anatomy
University of Helsinki, Pentti Somerharju / Principal Investigator
University of Helsinki, Department of Anatomy
University of Helsinki, Division of Pharmaceutical Chemistry and Technology
University of Helsinki, Medicum
Date: 2019-03
Language: eng
Number of pages: 12
Belongs to series: Biochimie
ISSN: 0300-9084
URI: http://hdl.handle.net/10138/299791
Abstract: ORP2 is a sterol-binding protein with documented functions in lipid and glucose metabolism, Akt signaling, steroidogenesis, cell adhesion, migration and proliferation. Here we investigate the interactions of ORP2 with phosphoinositides (PIPs) by surface plasmon resonance (SPR), its affinity for cholesterol with a pull-down assay, and its capacity to transfer sterol in vitro. Moreover, we determine the effects of wild-type (wt) ORP2 and a mutant with attenuated PIP binding, ORP2(mHHK), on the subcellular distribution of cholesterol, and analyze the interaction of ORP2 with the related cholesterol transporter ORP1L. ORP2 showed specific affinity for PI(4,5)P-2, PI(3,4,5)P-3 and PI(4)P, with suggestive K-d values in the mu M range. Also binding of cholesterol by ORP2 was detectable, but a K-d could not be determined. Wt ORP2 was in HeLa cells mainly detected in the cytosol, ER, late endosomes, and occasionally on lipid droplets (LDs), while ORP2(mHHK) displayed an enhanced LD localization. Overexpression of wt ORP2 shifted the D4H cholesterol probe away from endosomes, while ORP2(mHHK) caused endosomal accumulation of the probe. Although ORP2 failed to transfer dehydroergosterol in an in vitro assay where OSBP is active, its knock-down resulted in the accumulation of cholesterol in late endocytic compartments, as detected by both D4H and filipin probes. Interestingly, ORP2 was shown to interact and partially co-localize on late endosomes with ORP1L, a cholesterol transporter/sensor at ER-late endosome junctions. Our data demonstrates that ORP2 binds several phosphoinositides, both PI(4)P and multiply phosphorylated species. ORP2 regulates the subcellular distribution of cholesterol dependent on its PIP-binding capacity. The interaction of ORP2 with ORP1L suggests a concerted action of the two ORPs. (C) 2018 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.
Subject: Cholesterol
Lipid transfer
OSBP-related protein
Phosphoinositide
Surface plasmon resonance
OXYSTEROL-BINDING-PROTEIN
OSBP-RELATED PROTEIN-2
MEMBRANE CONTACT SITES
PHOSPHATIDYLSERINE TRANSPORT
PLASMA-MEMBRANE
LIPID DROPLETS
STEROL
ER
FAMILY
RAB7-RILP-P150(GLUED)
3111 Biomedicine
1182 Biochemistry, cell and molecular biology
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