Superparamagnetic Nanoparticles for Lysosome Isolation to Identify Spatial Alterations in Lysosomal Protein and Lipid Composition.


Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
18 12 2020
Historique:
entrez: 30 12 2020
pubmed: 31 12 2020
medline: 31 12 2020
Statut: epublish

Résumé

Lysosomes are dynamic organelles that serve as regulatory hubs in cellular homeostasis. Changes in lysosome morphology, composition, and turnover are typically linked to disease. These characteristics make enrichment protocols based on biophysical parameters challenging. However, organelle enrichment methods are essential to facilitate their biomolecular analysis. We describe the synthesis and use of superparamagnetic iron oxide nanoparticles (SPIONs) for high-yield purification of lysosomes compatible with "omics" analysis. NANOLYSE (Nanoparticles for Lysosome Isolation) provides a reliable strategy in fingerprinting the biomolecular composition of lysosomes. For complete details on the use and execution of this protocol, please refer to Tharkeshwar et al. (2017).

Identifiants

pubmed: 33377016
doi: 10.1016/j.xpro.2020.100122
pii: S2666-1667(20)30109-X
pmc: PMC7756972
doi:

Substances chimiques

Lipids 0
Proteins 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Pagination

100122

Informations de copyright

© 2020 The Authors.

Déclaration de conflit d'intérêts

The authors declare no competing interests.

Références

Cell. 2016 Jun 30;166(1):193-208
pubmed: 27293189
J Am Chem Soc. 2002 Jul 17;124(28):8204-5
pubmed: 12105897
Proteomics. 2018 Mar;18(5-6):e1700113
pubmed: 29125683
Methods Cell Biol. 2015;126:21-43
pubmed: 25665439
Nat Methods. 2012 Jul;9(7):671-5
pubmed: 22930834
Sci Rep. 2017 Jan 30;7:41408
pubmed: 28134274

Auteurs

Arun Kumar Tharkeshwar (AK)

Laboratory of Neurobiology, VIB-Center for Brain & Disease Research, Leuven, Belgium.
Department of Neurosciences, KU Leuven, Leuven, Belgium.

David Demedts (D)

Laboratory for Membrane Trafficking, VIB-Center for Brain and Disease Research, Leuven, Belgium.
Department of Neurosciences, KU Leuven, Leuven, Belgium.

Wim Annaert (W)

Laboratory for Membrane Trafficking, VIB-Center for Brain and Disease Research, Leuven, Belgium.
Department of Neurosciences, KU Leuven, Leuven, Belgium.

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Classifications MeSH