Metabolic labelling of choline phospholipids probes ABCA3 transport in lamellar bodies

Li, Yang; Kinting, Susanna; Höppner, Stefanie; Forstner, Maria; Uhl, Olaf; Koletzko, Berthold; Griese, Matthias

doi:10.1016/j.bbalip.2019.158516

Cited by 8 publications

(16 citation statements)

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“…Human ABCA3 is involved in the export of surfactant lipids-such as PC, PG, and cholesterol-from the cytosol into the intracellular vesicles (8,(11)(12)(13)(14). The two positively charged cavities within the cytosolic membrane leaflet of TMDs and the lipid-like densities observed within the two cavities strongly suggest that these two cavities are most likely the substrate binding sites, responsible for the export of surfactant lipid across the limiting membrane of LBs.…”

Section: Discussionmentioning

confidence: 99%

Cryo-EM structures of the human surfactant lipid transporter ABCA3

et al. 2022

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The adenosine 5′-triphosphate (ATP)–binding cassette (ABC) transporter ABCA3 plays a critical role in pulmonary surfactant biogenesis. Mutations in human ABCA3 have been recognized as the most frequent causes of inherited surfactant dysfunction disorders. Despite two decades of research, in vitro biochemical and structural studies of ABCA3 are still lacking. Here, we report the cryo-EM structures of human ABCA3 in two distinct conformations, both at resolution of 3.3 Å. In the absence of ATP, ABCA3 adopts a “lateral-opening” conformation with the lateral surfaces of transmembrane domains (TMDs) exposed to the membrane and features two positively charged cavities within the TMDs as potential substrate binding sites. ATP binding induces pronounced conformational changes, resulting in the collapse of the potential substrate binding cavities. Our results help to rationalize the disease-causing mutations in human ABCA3 and suggest a conserved “lateral access and extrusion” mechanism for both lipid export and import mediated by ABCA transporters.

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Section: Discussionmentioning

confidence: 99%

Cryo-EM structures of the human surfactant lipid transporter ABCA3

et al. 2022

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“…In terms of radiolabeled substrates, cholesterol is by far the most frequently used genuine ABCA1 substrate, 230 , 245 , 249 , 255 , 260 , 264 , 265 , 267 - 270 , 272 , 273 , 276 , 278 , 289 - 292 , 305 , 306 , 313 , 315 , 318 , 329 , 338 , 339 , 354 , 366 , 367 , 381 , 384 , 385 , 393 - 395 , 404 , 408 , 419 , 464 , 467 , 499 , 512 followed by phospholipid(-components). 249 , 255 , 267 , 269 , 273 , 311 , 464 , 467 , 514 However, other substrates have also been used. These substrates include mostly molecules with sterane scaffold, such as β-sitosterol (ABCA1) 262 and estradiol-β-glucuronide (ABCA8).…”

Section: Part Ii: Pipeline Development To Gain Novel Diagnostics and Therapeuticsmentioning

confidence: 99%

“… 136 Notably, radiolabeled substrates are very effective in terms of accurate tracing of protein function, as these molecules are not changed in their molecular integrity in contrast to fluorescence probes. 171 , 201 , 238 , 251 , 252 , 254 , 256 , 258 , 261 , 271 , 282 , 308 , 319 , 321 , 330 , 332 , 335 , 342 , 360 , 379 , 389 , 390 , 392 , 397 , 402 , 406 , 455 , 456 , 468 , 514 - 518 On the downside, conducting these experiments is constrained to regulatory requirements and requires extensive staff training as well as expensive safety measures and laboratory equipment.…”

Section: Part Ii: Pipeline Development To Gain Novel Diagnostics and Therapeuticsmentioning

confidence: 99%

“… 271 , 282 , 319 , 321 , 330 , 332 , 455 , 456 , 515 - 517 Other fluorophore-labeled dyes have been reported, too, including the sterane analog dansylestramustine, 171 , 238 , 518 and propargyl choline, which is processed in vitro into propargylated phospholipids. 514 …”

Section: Part Ii: Pipeline Development To Gain Novel Diagnostics and Therapeuticsmentioning

confidence: 99%

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Strategies to gain novel Alzheimer’s disease diagnostics and therapeutics using modulators of ABCA transporters

Pahnke

Bascuñana

Brackhan

et al. 2021

Free Neuropathology

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Adenosine-triphosphate-(ATP)-binding cassette (ABC) transport proteins are ubiquitously present membrane-bound efflux pumps that distribute endo- and xenobiotics across intra- and intercellular barriers. Discovered over 40 years ago, ABC transporters have been identified as key players in various human diseases, such as multidrug-resistant cancer and atherosclerosis, but also neurodegenerative diseases, such as Alzheimer’s disease (AD). Most prominent and well-studied are ABCB1, ABCC1, and ABCG2, not only due to their contribution to the multidrug resistance (MDR) phenotype in cancer, but also due to their contribution to AD. However, our understanding of other ABC transporters is limited, and most of the 49 human ABC transporters have been largely neglected as potential targets for novel small-molecule drugs. This is especially true for the ABCA subfamily, which contains several members known to play a role in AD initiation and progression. This review provides up-to-date information on the proposed functional background and pathological role of ABCA transporters in AD. We also provide an overview of small-molecules shown to interact with ABCA transporters as well as potential in silico , in vitro , and in vivo methodologies to gain novel templates for the development of innovative ABC transporter-targeting diagnostics and therapeutics.

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“…Recently, Griese and co-workers employed PC labeling using PCho to study the activity of ATP-binding cassette sub-family A member 3 (ABCA3) in the formation and trafficking of lipids into the lamellar body (LB), processes that are dysregulated in chronic interstitial lung diseases (Li et al, 2019b). This labeling protocol was first validated for the fluorescence imaging of PC lipid products in ABCA3+ vesicles, with all chosen vesicles labeled after 24 hours.…”

Section: Investigations Of Lipid Metabolism Trafficking Localization and Protein Bindingmentioning

confidence: 99%

Metabolic labeling of glycerophospholipids via clickable analogs derivatized at the lipid headgroup

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2020

Chemistry and Physics of Lipids

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Metabolic labeling, in which substrate analogs containing diminutive tags can infiltrate biosynthetic pathways and generate labeled products in cells, has led to dramatic advancements in the means by which complex biomolecules can be detected and biological processes can be elucidated. Within this realm, metabolic labeling of lipid products, particularly in a manner that is headgroup-specific, brings about a number of technical challenges including the complexity of lipid metabolic pathways as well as the simplicity of biosynthetic precursors to headgroup functionality. As such, only a handful of strategies for metabolic labeling of lipids have thus far been reported. However, these approaches provide enticing examples of how strategic modifications to substrate structures, particularly by introducing clickable moieties, can enable the hijacking of lipid biosynthesis. Furthermore, early work in this field has led to an explosion in diverse applications by which these techniques have been exploited to answer key biological questions or detect and track various lipid-containing biological entities. In this article, we review these efforts and emphasize recent advancements in the development and application of lipid metabolic labeling strategies.

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Metabolic labelling of choline phospholipids probes ABCA3 transport in lamellar bodies

Cited by 8 publications

References 41 publications

Cryo-EM structures of the human surfactant lipid transporter ABCA3

Cryo-EM structures of the human surfactant lipid transporter ABCA3

Strategies to gain novel Alzheimer’s disease diagnostics and therapeutics using modulators of ABCA transporters

Metabolic labeling of glycerophospholipids via clickable analogs derivatized at the lipid headgroup

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