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Mouse osteoclast secretory lysosomes nLC-ESI-MS/MS

ABSTRACT: To survey the proteome of osteoclast secretory lysosomes, we used superparamagnetic iron oxide nanoparticles (SPIONs) to enrich for these endo-lysosomal-related organelles from murine osteoclast cultures. Briefly, large scale murine bone marrow monocyte (BMM)-derived osteoclast cultures were ‘pulsed’ with SPIONs to encourage uptake into endosomes and then ‘chased’ into secretory lysosomes upon the convergence of SPION-loaded endosomes with lysosomes and secretory pathways. Following the ‘pulse-chase’, osteoclasts were homogenized, SPION-loaded organelles captured-from post-nuclear supernatants using magnetic columns, and enriched organelles eluted and processed for 1D in-gel digestion and mass spectrometry.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Bone Marrow, Macrophage

SUBMITTER: Amy Ribet

LAB HEAD: Nathan Pavlos

PROVIDER: PXD037006 | Pride | 2023-01-25

REPOSITORIES: Pride

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			Action	DRS
	AR_20191031_Biological_Triplicate_WT_homog_vs_lyso_pooled_normalised.msf	Msf
	AR_20220405_Biological_Triplicate_WT_homog_vs_lyso_pooled_normalised.mgf	Mgf
	Homogenate_BioRep1_WT_Fraction1.raw	Raw
	Homogenate_BioRep1_WT_Fraction10.raw	Raw
	Homogenate_BioRep1_WT_Fraction2.raw	Raw

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