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Identification and Partial Characterization of Two Populations of Prostasomes by a Combination of Dynamic Light Scattering and Proteomic Analysis

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Abstract

Prostasomes are vesicles secreted by prostate epithelial cells and are found in abundance in the semen. Here we characterized two different prostasome populations isolated from human seminal fluid. Prostasomes were isolated using differential centrifugation, while dynamic light scattering (DLS) was used to characterize their size and size distribution. Their protein content was analyzed using two-dimensional electrophoresis and mass spectrometry. DLS showed two distinct prostasome subpopulations in centrifuged seminal plasma, with an average hydrodynamic radius of 80 and 300 nm. The larger population was isolated after centrifugation at 20,000×g (P20), while the smaller one was recovered at 100,000×g (P100). The two fractions had a similar lipid composition, showing an elevated content of sphingomyelin and cholesterol. The P100 vesicles showed a significant over-expression of proteins involved in the endosomal sorting complexes required for transport (ESCRT) machinery such as Alix, TSG101, and syntenin-1. Some proteins possibly involved in prostate cancer were present only in one specific population (TMPRSS2 in P100 and VCP in P20). The different size and protein profile in the two subpopulations of prostasomes might support differential roles of the semen vesicles toward the target cells, and/or different secretion pathways from the organ of origin.

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Acknowledgments

This work was funded by Fondazione Cassa di Risparmio di Perugia (Grant Number 2010.011.0396).

Conflicts of interest

The authors declare that there are no conflicts of interest.

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    Authors and Affiliations

    1. Sezione di Neurologia, Dipartimento di Medicina, Università degli Studi di Perugia, Perugia, Italy

      Davide Chiasserini

    2. Dipartimento di Scienze Agrarie, Alimentari ed Ambientali, Unità di Ricerca di Biochimica e Biologia Molecolare, Via del Giochetto, 06125, Perugia, Italy

      Michela Mazzoni & Carlo Alberto Palmerini

    3. Sez di Fisiologia e Biochimica, Dipartimento di Medicina Sperimentale, Università degli Studi di Perugia, Perugia, Italy

      Federica Susta, Pier Luigi Orvietani & Luciano Binaglia

    4. Dipartimento di Fisica and CNR-IPCF, Sapienza Università di Roma, piazzale A. Moro, 2, 00185, Rome, Italy

      Federico Bordi

    5. Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Viale Regina Elena, 291, 00161, Rome, Italy

      Federico Bordi

    6. ISC-CNR, UOS Sapienza, P.le Aldo Moro 2, 00185, Rome, Italy

      Simona Sennato

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    Davide Chiasserini and Michela Mazzoni have contributed equally to the work.

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    Chiasserini, D., Mazzoni, M., Bordi, F. et al. Identification and Partial Characterization of Two Populations of Prostasomes by a Combination of Dynamic Light Scattering and Proteomic Analysis. J Membrane Biol 248, 991–1004 (2015). https://doi.org/10.1007/s00232-015-9810-0

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