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

Identification and Partial Characterization of Two Populations of Prostasomes by a Combination of... 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Journal of Membrane Biology Springer Journals

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

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Publisher
Springer US
Copyright
Copyright © 2015 by Springer Science+Business Media New York
Subject
Life Sciences; Biochemistry, general; Human Physiology
ISSN
0022-2631
eISSN
1432-1424
D.O.I.
10.1007/s00232-015-9810-0
Publisher site
See Article on Publisher Site

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.

Journal

The Journal of Membrane BiologySpringer Journals

Published: Jun 12, 2015

References

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