Reversible chemical dimerizer-induced recovery of PIP2 levels moves clathrin to the plasma membrane

Martina Schifferer; Suihan Feng; Frank Stein; Christian Tischer; Carsten Schultz

doi:10.1016/j.bmc.2015.03.048

Reversible chemical dimerizer-induced recovery of PIP₂ levels moves clathrin to the plasma membrane

Martina Schifferer, Suihan Feng, Frank Stein, Christian Tischer, Carsten Schultz

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

Chemical dimerizers are powerful non-invasive tools for bringing molecules together inside intact cells. We recently introduced a rapidly reversible chemical dimerizer system which enables transient translocation of enzymes to and from the plasma membrane (PM). Here we have applied this system to transiently activate phosphatidylinositol 4,5-bisphosphate (PIP₂) breakdown at the PM via translocation of phosphoinositide 5-phosphatase (5Ptase). We found that the PIP₂ sensor phospholipase C-δ PH domain (PLCδ-PH) is released from the PM upon addition of the reversible chemical dimerizer rCD1. By outcompeting rCD1, rapid release of the 5Ptase from the PM is followed by PIP₂ recovery. This permits the observation of the PIP₂-dependent clathrin assembly at the PM.

Original language	English (US)
Pages (from-to)	2862-2867
Number of pages	6
Journal	Bioorganic and Medicinal Chemistry
Volume	23
Issue number	12
DOIs	https://doi.org/10.1016/j.bmc.2015.03.048
State	Published - May 29 2015
Externally published	Yes

Keywords

Adapter proteins
Clathrin
Endocytosis
Phosphatidylinositol 4,5-bisphosphate
Phosphoinositol 5-phosphatase

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmaceutical Science
Drug Discovery
Clinical Biochemistry
Organic Chemistry

Access to Document

10.1016/j.bmc.2015.03.048

Cite this

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title = "Reversible chemical dimerizer-induced recovery of PIP2 levels moves clathrin to the plasma membrane",

abstract = "Chemical dimerizers are powerful non-invasive tools for bringing molecules together inside intact cells. We recently introduced a rapidly reversible chemical dimerizer system which enables transient translocation of enzymes to and from the plasma membrane (PM). Here we have applied this system to transiently activate phosphatidylinositol 4,5-bisphosphate (PIP2) breakdown at the PM via translocation of phosphoinositide 5-phosphatase (5Ptase). We found that the PIP2 sensor phospholipase C-δ PH domain (PLCδ-PH) is released from the PM upon addition of the reversible chemical dimerizer rCD1. By outcompeting rCD1, rapid release of the 5Ptase from the PM is followed by PIP2 recovery. This permits the observation of the PIP2-dependent clathrin assembly at the PM.",

keywords = "Adapter proteins, Clathrin, Endocytosis, Phosphatidylinositol 4,5-bisphosphate, Phosphoinositol 5-phosphatase",

author = "Martina Schifferer and Suihan Feng and Frank Stein and Christian Tischer and Carsten Schultz",

note = "Funding Information: M.S. is a fellow of the EMBL Interdisciplinary Postdoc (EIPOD) Programme, co-funded by the EU. This work was supported by the DFG (Transregio 83). We are grateful for technical support of EMBL{\textquoteright}s Advanced Light Microscopy Facility (ALMF). We thank Ori Avinoam (EMBL) for helping with the clathrin experiments and Madeleine Schultz (EMBL) for critical reading of the manuscript. Publisher Copyright: {\textcopyright} 2015 Published by Elsevier Ltd.",

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AU - Feng, Suihan

AU - Stein, Frank

AU - Tischer, Christian

AU - Schultz, Carsten

N1 - Funding Information: M.S. is a fellow of the EMBL Interdisciplinary Postdoc (EIPOD) Programme, co-funded by the EU. This work was supported by the DFG (Transregio 83). We are grateful for technical support of EMBL’s Advanced Light Microscopy Facility (ALMF). We thank Ori Avinoam (EMBL) for helping with the clathrin experiments and Madeleine Schultz (EMBL) for critical reading of the manuscript. Publisher Copyright: © 2015 Published by Elsevier Ltd.

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N2 - Chemical dimerizers are powerful non-invasive tools for bringing molecules together inside intact cells. We recently introduced a rapidly reversible chemical dimerizer system which enables transient translocation of enzymes to and from the plasma membrane (PM). Here we have applied this system to transiently activate phosphatidylinositol 4,5-bisphosphate (PIP2) breakdown at the PM via translocation of phosphoinositide 5-phosphatase (5Ptase). We found that the PIP2 sensor phospholipase C-δ PH domain (PLCδ-PH) is released from the PM upon addition of the reversible chemical dimerizer rCD1. By outcompeting rCD1, rapid release of the 5Ptase from the PM is followed by PIP2 recovery. This permits the observation of the PIP2-dependent clathrin assembly at the PM.

AB - Chemical dimerizers are powerful non-invasive tools for bringing molecules together inside intact cells. We recently introduced a rapidly reversible chemical dimerizer system which enables transient translocation of enzymes to and from the plasma membrane (PM). Here we have applied this system to transiently activate phosphatidylinositol 4,5-bisphosphate (PIP2) breakdown at the PM via translocation of phosphoinositide 5-phosphatase (5Ptase). We found that the PIP2 sensor phospholipase C-δ PH domain (PLCδ-PH) is released from the PM upon addition of the reversible chemical dimerizer rCD1. By outcompeting rCD1, rapid release of the 5Ptase from the PM is followed by PIP2 recovery. This permits the observation of the PIP2-dependent clathrin assembly at the PM.

KW - Adapter proteins

KW - Clathrin

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KW - Phosphatidylinositol 4,5-bisphosphate

KW - Phosphoinositol 5-phosphatase

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