The proteome of extracellular vesicles released by clastic cells differs based on their substrate

ECU Author/Contributor (non-ECU co-authors, if there are any, appear on document)

Wellington J. Jr.,Chamberlain,Casey A.,Emory-Carter,Alyssa K. Rody (Creator)

Institution

East Carolina University (ECU )

Web Site: http://www.ecu.edu/lib/

Abstract: Extracellular vesicles (EVs) from osteoclasts are important regulators in intercellular communication. Here, we investigated the proteome of EVs from clastic cells plated on plastic(clasts), bone (osteoclasts) and dentin (odontoclasts) by two-dimensional high performanceliquid chromatography mass spectrometry seeking differences attributable to distinct mineralized matrices. A total of 1,952 proteins were identified. Of the 500 most abundant proteinsin EVs, osteoclast and odontoclast EVs were 83.3% identical, while clasts shared 70.7% ofthe proteins with osteoclasts and 74.2% of proteins with odontoclasts. For each protein, thedifferences between the total ion count values were mapped to an expression ratio histogram (Z-score) in order to detect proteins differentially expressed. Stabilin-1 and macrophage mannose receptor-1 were significantly-enriched in EVs from odontoclasts comparedwith osteoclasts (Z = 2.45, Z = 3.34) and clasts (Z = 13.86, Z = 1.81) and were abundant inodontoclast EVs. Numerous less abundant proteins were differentially-enriched. Subunits ofknown protein complexes were abundant in clastic EVs, and were present at levels consistent with them being in assembled protein complexes. These included the proteasome,COP1, COP9, the T complex and a novel sub-complex of vacuolar H+-ATPase (V-ATPase),which included the (pro) renin receptor. The (pro) renin receptor was immunoprecipitatedusing an anti-E-subunit antibody from detergent-solubilized EVs, supporting the idea thatthe V-ATPase subunits present were in the same protein complex. We conclude that theprotein composition of EVs released by clastic cells changes based on the substrate. ClasticEVs are enriched in various protein complexes including a previously undescribed VATPase sub-complex.

Additional Information

Publication

Other

Language: English

Date: 2019

Email this document to