Phosphoinositide profiling in complex lipid mixtures using electrospray ionization mass spectrometry

Wenk, Markus R.; Lucast, Louise; Paolo, Gilbert Di; Romanelli, Anthony J.; Suchy, Sharon F.; Nussbaum, Robert L.; Cline, Gary W.; Shulman, Gerald I.; McMurray, Walter J.; Camilli, Pietro De

doi:10.1038/nbt837

Cited by 244 publications

(219 citation statements)

References 19 publications

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“…phosphatidylserine (PS) are more abundant with 8.5% of total lipids (Wenk, Lucast et al 2003;Lemmon 2008). This emphasizes the fundamental importance and regulatory role of PIs in processes such as membrane trafficking and signaling,…”

Section: Phosphoinositides Determine Membrane Identitymentioning

confidence: 99%

A phosphoinositide conversion mechanism for exit from endosomes

Ketel¹,

Krauß²,

Nicot³

et al. 2016

Nature

165

177

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I owe special thanks to Marnix Wieffer and Michael Krauß, who spent a lot of time helping me to get started in the lab, to overcome numerous experimental problem, I was not sure how to deal with, and frequently joined in discussions about my project with excellent advice. I am really grateful for your experimental support, Michael, and the time you invested in the project during our paper revision.Further, I would like to thank Jocelyn Laporte and his group, especially Anne-Sophie Nicot, at the IGBMC in Strasbourg for their support and a very fruitful collaboration, and Carsten Schultz and his group at the EMBL in Heidelberg for their support and constant supply with PIP/AMs. I owe special thanks to Dmytro Puchkov for the ultrastructural analysis and Eberhard Krause for mass spectrometry done within this project. I also need to acknowledge Markus Wenk and Federico Torta at the Singapore Lipidomics Incubator for joining the project at a very late stage, when we urgently needed help from lipidomics specialists. It was a pity that experiments did not work out as planned.I would further like to express my gratitude to two very skilled technicians in the AG Haucke lab: Silke Zillmann and Delia Löwe. Without your help it would have been impossible to achieve so much within the limited amount of time I had during the paper revision. by VPS34-IN1 treatment................................................... 52 2.3.11 Fluorescence microscopy................................................................................. 53 2.3.12 Flow cytometry............................................................................................ III SummaryPhosphoinositides (PIs) are a minor class of short-lived phospholipids that serve as crucial signposts of membrane identity. Thereby, PIs full fill important functions in cell signaling and membrane transport. PI 4-phosphates such as phosphatitylinositol-4-phosphate (PI(4)P) and phosphatitylinositol-4,5-bisphosphate (PI(4,5)P 2 ) are enriched at the plasma membrane (PM), on secretory organelles and lysosomes, while PI 3-phosphates, i.e.phosphatitylinositol-3-phosphate (PI(3)P), are a hallmark of the endosomal system.Directional transport between these compartments, thus, requires regulated PI conversion.However, PI conversion in exit from PI(3)P-enriched endosomes en route to the PI(4)P-and PI(4,5)P 2 -positive PM in endosomal recycling remained unknown.Here, we report that cargo exit from endosomes requires removal of PI(3)P by the PI(3)P 3-phosphatase myotubularin 1 (MTM1), and concomitant PI(4)P synthesis by PI 4-kinase type II α (PI4K2α). Loss of MTM1 causes endosomal accumulation of PI(3)P and PI(3)P effector proteins, i.e. sorting nexins, Kif16b-mediated outward traffic of PI(3)P containing endosomes and sub-plasmalemmal accumulation of exocytosis-deficient endosomes. As PI4K2α associates with MTM1 and thereby facilitates membrane recruitment of MTM1, these phenotypic changes are mimicked by loss of PI4K2α. The conversion of PI(3)P-to-PI(4)P is paralleled by a switch i...

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Section: Phosphoinositides Determine Membrane Identitymentioning

confidence: 99%

A phosphoinositide conversion mechanism for exit from endosomes

Ketel¹,

Krauß²,

Nicot³

et al. 2016

Nature

165

177

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“…The deficiency of ocrl1 leads to elevated cellular levels of PIP 2 (Wenk et al 2003;Zhang et al 1998). However, it is unknown at present why a deficiency of this widely expressed protein primarily affects the lens, kidney and brain in Lowe syndrome.…”

Section: Introductionmentioning

confidence: 99%

Abnormal bradykinin signalling in fibroblasts deficient in the PIP₂ 5‐phosphatase, ocrl1

Suchy

Cronin

Nussbaum

2009

J of Inher Metab Disea

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Summary The oculocerebrorenal syndrome of Lowe (Lowe syndrome) is an X‐linked disorder of phosphatidylinositol metabolism characterized by congenital cataracts, renal proximal tubulopathy and neurological deficits. The disorder is due to the deficiency of the phosphatidylinositol 4,5‐bisphosphate (PIP2) 5‐phosphatase, ocrl1. PIP2 is critical for numerous cellular processes, including cell signalling, actin reorganization and protein trafficking, and is chronically elevated in patients with Lowe syndrome. The elevation of PIP2 cells of patients with Lowe syndrome provides the unique opportunity to investigate the roles of this phospholipid in fundamental cellular processes. We previously demonstrated that ocrl1 deficiency causes alterations in the actin cytoskeleton. Since actin remodelling is strongly activated by [Ca+2], which increases in response to IP3 production, we hypothesized that altered calcium signalling might contribute to the observed abnormalities in actin organization. Here we report a specific increase in bradykinin‐induced Ca+2 mobilization in Lowe fibroblasts. We show that the abnormal bradykinin signalling occurs in spite of normal total cellular receptor content. These data point to a novel role for ocrl1 in agonist‐induced calcium release.

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“…1). PIP2, along with phosphatidylinositol (PI) and phosphatidylinositol 4-phosphate (PI-4-P) make up the bulk of inositide lipids in vivo [2,3]. The biological importance of PIP2 and PIP3 is exemplified by the effect of mutations in the proteins that convert the former into the latter.…”

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confidence: 99%

“…This has farreaching consequences, as PI is the commonest inositide, forming an important fraction of animal [2,3,24], plant [25], and some bacterial [26] membranes. This understanding of the physical behaviour of PI has recently been shown in a new light by a recent report from Atilla-Gokcumen et al about the lipid composition of HeLa cells [24].…”

mentioning

confidence: 99%

“…1 The structure of the inositide lipid group. All combinations and permutations of phosphates at R 1 , R 2 and R 3 have been found in vivo, with phosphatidylinositol-4,5-bisphosphate (PI-4,5-P 2 ; R 2 and R 3 = -PO 3 H − , R 1 = H), phosphatidylinositol 4-phosphate (PI-4-P; R 2 = -PO 3 H − , R 1 and R 3 = H), and phosphatidylinositol (PI, R 1 , R 2 and R 3 = H) forming the bulk of the mass of inositides under typical physiological conditions [2,3] …”

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The physical influence of inositides—a disproportionate effect?

Furse

2014

J Chem Biol

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After the initial observation that lipids form a considerable part of biological membranes, the details of the physical role of lipids in biological systems have emerged gradually. There have been few 'Eureka' moments in which a class or individual lipid has appeared as a game-changing physical player. However, evidence collected in the last five years suggests that that notion may be about to change. In chemical biology studies, inositides are increasingly showing themselves to be lipids that have a physical influence on membrane systems that is as strong as their biological (signalling) one. Additionally, recent evidence has shown that the concentration of at least one inositide changes during important stages of the cell cycle, and not in a manner consistent with its traditional signalling roles. The balance between these data is explored and a forward-looking view is proposed. The apparent ubiquity of these two inositides in mammalian systems has aroused research interest from several angles. There is a mounting body of evidence that it is not only mammals that make use of PIP2 as a signalling molecule, but also insects [9] and plants [10]. Several studies have suggested that PIP2 has a physical role that might be quite separate from its biological one. The particular processes in which PIP2 is a protagonist include membrane trafficking [11,12], vesicle formation [12,13], clatharin-mediated endocytosis [13,14], and in membrane dynamics [15]. There is also some indication that the activity of phospholipase Cγ, for which PIP2 is a substrate, may have a physical role in membrane fusion [16]. It also seems likely that PIP2 is a team player, acting with other lipids such that certain physical processes may be completed. A study of pollen tubes in Arabidopsis provided evidence that a particular balance between PIP2 and PI-4-P is required for endocytosis to be successful [17]. There have not yet been any comprehensive studies of the phase behaviour of PIP2, and thus, it is not yet clear whether it is typically a bilayer-forming lipid or one that has an energetic preference for a curved topology.Despite the paucity of phase behaviour data on PIP2, the phase behaviour of the physiological precursor to PIP2, PI-4-P, has been researched. Furse et al. reported that PI-4-P is able to induce an inverse hexagonal phase in the typically-lamellar PC, at 2 mol % [18], strongly suggesting that PI-4-P is not a

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Phosphoinositide profiling in complex lipid mixtures using electrospray ionization mass spectrometry

Cited by 244 publications

References 19 publications

A phosphoinositide conversion mechanism for exit from endosomes

A phosphoinositide conversion mechanism for exit from endosomes

Abnormal bradykinin signalling in fibroblasts deficient in the PIP₂ 5‐phosphatase, ocrl1

The physical influence of inositides—a disproportionate effect?

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Phosphoinositide profiling in complex lipid mixtures using electrospray ionization mass spectrometry

Cited by 244 publications

References 19 publications

A phosphoinositide conversion mechanism for exit from endosomes

A phosphoinositide conversion mechanism for exit from endosomes

Abnormal bradykinin signalling in fibroblasts deficient in the PIP2 5‐phosphatase, ocrl1

The physical influence of inositides—a disproportionate effect?

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Abnormal bradykinin signalling in fibroblasts deficient in the PIP₂ 5‐phosphatase, ocrl1