Mouse osteoclast homogenates nLC-ESI-MS/MS - WT vs Slc37a2 KO

ABSTRACT: To survey the proteomic differences between WT and Slc37a2 knockout osteoclasts at the whole cell and secretory lysosome levels, 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 as well as homogenates eluted and processed for 1D in-gel digestion and mass spectrometry. The samples presented here correspond to the proteome of WT and Slc37a2 knockout homogenates with secretory lysosome proteomes have been shared in another submission.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Bone Marrow

SUBMITTER: Amy Ribet  

LAB HEAD: Nathan Pavlos

PROVIDER: PXD037910 | Pride | 2023-01-25

REPOSITORIES: Pride