Greasing the Wheels of Lipid Biology with Chemical Tools

Bumpus, Timothy W.; Baskin, Jeremy M.

doi:10.1016/j.tibs.2018.09.011

Cited by 61 publications

(65 citation statements)

References 82 publications

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“…Another factor key to the development of optoPLD was IMPACT, a chemical biology tool that we had originally designed to study the activity and localization of endogenous mammalian PLDs ( Bumpus and Baskin, 2016 , 2017 , 2018 ). Here, we confirmed that IMPACT could be applied to quantify the activity of an exogenous, bacterial PLD within mammalian cells.…”

Section: Discussionmentioning

confidence: 99%

“…In summary, optoPLD is a tool for precise, spatiotemporally controlled production of local pools of PA on individual organelle membranes that should complement existing toolsets for modulating PA using inhibition or knockout/knockdown of biosynthetic enzymes. OptoPLD interfaces with a variety of imaging-based tools for monitoring the dynamics of PA metabolism, including genetically encoded biosensors, FRET-based imaging of PLD–substrate binding events, and our recently developed chemoenzymatic method, IMPACT ( Zhang et al, 2014 ; Kassas et al, 2012 , 2017 ; Bumpus and Baskin, 2018 ; Kim et al, 2015 ; Lu et al, 2016 ; Hughes et al, 2002 ). We validated the physiological relevance of optoPLD-derived PA pools to confirm and elucidate differential roles of PA signaling in the modulation of Hippo signaling.…”

Section: Discussionmentioning

confidence: 99%

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Spatiotemporal control of phosphatidic acid signaling with optogenetic, engineered phospholipase Ds

Tei

Baskin

2020

Journal of Cell Biology

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Phosphatidic acid (PA) is both a central phospholipid biosynthetic intermediate and a multifunctional lipid second messenger produced at several discrete subcellular locations. Organelle-specific PA pools are believed to play distinct physiological roles, but tools with high spatiotemporal control are lacking for unraveling these pleiotropic functions. Here, we present an approach to precisely generate PA on demand on specific organelle membranes. We exploited a microbial phospholipase D (PLD), which produces PA by phosphatidylcholine hydrolysis, and the CRY2–CIBN light-mediated heterodimerization system to create an optogenetic PLD (optoPLD). Directed evolution of PLD using yeast membrane display and IMPACT, a chemoenzymatic method for visualizing cellular PLD activity, yielded a panel of optoPLDs whose range of catalytic activities enables mimicry of endogenous, physiological PLD signaling. Finally, we applied optoPLD to elucidate that plasma membrane, but not intracellular, pools of PA can attenuate the oncogenic Hippo signaling pathway. OptoPLD represents a powerful and precise approach for revealing spatiotemporally defined physiological functions of PA.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Spatiotemporal control of phosphatidic acid signaling with optogenetic, engineered phospholipase Ds

Tei

Baskin

2020

Journal of Cell Biology

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“…Localized production or accumulation of intracellular signaling agents is a key feature of signal transduction pathways. Chemical imaging tools have helped pave the way to understand biological signaling pathways dependent on second messengers (2)(3)(4)66). We initiated this study with the goal of establishing an activity-based imaging tool to enable precise monitoring of the production of the lipid second messenger PA by PLD enzymes.…”

Section: Discussionmentioning

confidence: 99%

A real-time, click chemistry imaging approach reveals stimulus-specific subcellular locations of phospholipase D activity

Liang

Tei

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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The fidelity of signal transduction requires spatiotemporal control of the production of signaling agents. Phosphatidic acid (PA) is a pleiotropic lipid second messenger whose modes of action differ based on upstream stimulus, biosynthetic source, and site of production. How cells regulate the local production of PA to effect diverse signaling outcomes remains elusive. Unlike other second messengers, sites of PA biosynthesis cannot be accurately visualized with subcellular precision. Here, we describe a rapid, chemoenzymatic approach for imaging physiological PA production by phospholipase D (PLD) enzymes. Our method capitalizes on the remarkable discovery that bulky, hydrophilic trans-cyclooctene–containing primary alcohols can supplant water as the nucleophile in the PLD active site in a transphosphatidylation reaction of PLD’s lipid substrate, phosphatidylcholine. The resultant trans-cyclooctene–containing lipids are tagged with a fluorogenic tetrazine reagent via a no-rinse, inverse electron-demand Diels–Alder (IEDDA) reaction, enabling their immediate visualization by confocal microscopy in real time. Strikingly, the fluorescent reporter lipids initially produced at the plasma membrane (PM) induced by phorbol ester stimulation of PLD were rapidly internalized via apparent nonvesicular pathways rather than endocytosis, suggesting applications of this activity-based imaging toolset for probing mechanisms of intracellular phospholipid transport. By instead focusing on the initial 10 s of the IEDDA reaction, we precisely pinpointed the subcellular locations of endogenous PLD activity as elicited by physiological agonists of G protein-coupled receptor and receptor tyrosine kinase signaling. These tools hold promise to shed light on both lipid trafficking pathways and physiological and pathological effects of localized PLD signaling.

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“…Lipids are hydrophobic biomolecules that can be broadly divided by their chemical composition into three main categories: simple lipids (eg, triglycerides, TGs), compound lipids (eg, sphingolipids) and derived lipids (eg, cholesterol). 18 However, most lipids are stored in adipose tissue in the form of the simple lipid TG. There are two main sources of lipids in the body: dietary intake and in vivo synthesis.…”

Section: Mechanisms By Which Pcbs Affect Hepatic Lipid Accumulationmentioning

confidence: 99%

<p>Understanding the Multiple Effects of PCBs on Lipid Metabolism</p>

Shan

Chen

et al. 2020

DMSO

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Polychlorinated biphenyls (PCBs) are a typical class of environmental contaminants recently shown to be metabolism-disrupting chemicals. Lipids are a highly complex group of biomolecules that not only form the structural basis of biofilms but also act as signaling molecules and energy sources. Lipid metabolic disorders contribute to multiple diseases, including obesity, diabetes, fatty liver, and metabolic syndromes. Although previous literature has reported that PCBs can affect lipid metabolism, including lipid synthesis, uptake, and elimination, few systematic summaries of the detailed process of lipid metabolism caused by PCB exposure have been published. Lipid metabolic processes involve many molecules; however, the key factors that are sensitive to PCB exposure have not been fully clarified. Here, we summarize the recent developments in PCB research with a focus on biomarkers of lipid metabolic disorders related to environmental exposures.

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Greasing the Wheels of Lipid Biology with Chemical Tools

Cited by 61 publications

References 82 publications

Spatiotemporal control of phosphatidic acid signaling with optogenetic, engineered phospholipase Ds

Spatiotemporal control of phosphatidic acid signaling with optogenetic, engineered phospholipase Ds

A real-time, click chemistry imaging approach reveals stimulus-specific subcellular locations of phospholipase D activity

<p>Understanding the Multiple Effects of PCBs on Lipid Metabolism</p>

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