Dataset Information

Systematic comparison of strategies for the enrichment of lysosomes by data independent acquisition

ABSTRACT: We describe a comparison of individual approaches for the enrichment of lysosomes comparing four different strategies using data independent acquisition (DIA). As a benchmark, we chose a crude enrichment strategy, which is the centrifugation of the PNS at 20,000 x g termed organelle enriched pellet (OEP), a two-step density gradient centrifugation (SDGC) approach employing sucrose, histodenz, and percoll, the SPIONs approach, which is based on the endocytosis of dextran coated nano-particles and their delivery to the lysosomal compartment, and the TMEM192-HA immunoprecipitation a 3 x HA tagged version of the lysosomal membrane protein TMEM192 used for lysosome isolation by anti-HA antibodies immobilized on magnetic beads. We evaluated the abundance and distributions of lysosomal, as well total proteins, allowing for the determination of lysosomal enrichment among the different strategies.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Early Embryonic Cell

SUBMITTER: Jasjot Singh

LAB HEAD: Dominic Winter

PROVIDER: PXD015098 | Pride | 2020-01-21

REPOSITORIES: Pride

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Publications

Systematic Comparison of Strategies for the Enrichment of Lysosomes by Data Independent Acquisition.

Singh Jasjot J Kaade Edgar E Muntel Jan J Bruderer Roland R Reiter Lukas L Thelen Melanie M Winter Dominic D

Journal of proteome research 20191210 1

In mammalian cells, the lysosome is the main organelle for the degradation of macromolecules and the recycling of their building blocks. Correct lysosomal function is essential, and mutations in every known lysosomal hydrolase result in so-called lysosomal storage disorders, a group of rare and often fatal inherited diseases. Furthermore, it is becoming more and more apparent that lysosomes play also decisive roles in other diseases, such as cancer and common neurodegenerative disorders. This le ...[more]

PMID: 31738065

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Dataset Information

Systematic comparison of strategies for the enrichment of lysosomes by data independent acquisition

Publications

Systematic Comparison of Strategies for the Enrichment of Lysosomes by Data Independent Acquisition.

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