The Ubiquitin-Proteasome System Participates in Sperm Surface Subproteome Remodeling during Boar Sperm Capacitation
Sperm capacitation is a complex process endowing biological and biochemical changes to a spermatozoon for a successful encounter with an oocyte. The present study focused on the role of the ubiquitin–proteasome system (UPS) in the remodeling of the sperm surface subproteome. The sperm surface subpro...
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MDPI AG
2023-06-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/13/6/996 |
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| author | Michal Zigo Karl Kerns Peter Sutovsky |
| author_facet | Michal Zigo Karl Kerns Peter Sutovsky |
| author_sort | Michal Zigo |
| collection | DOAJ |
| description | Sperm capacitation is a complex process endowing biological and biochemical changes to a spermatozoon for a successful encounter with an oocyte. The present study focused on the role of the ubiquitin–proteasome system (UPS) in the remodeling of the sperm surface subproteome. The sperm surface subproteome from non-capacitated and in vitro capacitated (IVC) porcine spermatozoa, with and without proteasomal inhibition, was selectively isolated. The purified sperm surface subproteome was analyzed using high-resolution, quantitative liquid chromatography–mass spectrometry (LC-MS) in four replicates. We identified 1680 HUGO annotated proteins, out of which we found 91 to be at least 1.5× less abundant (<i>p</i> < 0.05) and 141 to be at least 1.5× more abundant (<i>p</i> < 0.05) on the surface of IVC spermatozoa. These proteins were associated with sperm capacitation, hyperactivation, metabolism, acrosomal exocytosis, and fertilization. Abundances of 14 proteins were found to be significantly different (<i>p</i> < 0.05), exceeding a 1.5-fold abundance between the proteasomally inhibited (100 µM MG132) and vehicle control (0.2% ethanol) groups. The proteins NIF3L1, CSE1L, NDUFB7, PGLS, PPP4C, STK39, and TPRG1L were found to be more abundant; while BPHL, GSN, GSPT1, PFDN4, STYXL1, TIMM10, and UBXN4 were found to be less abundant in proteasomally inhibited IVC spermatozoa. Despite the UPS having a narrow range of targets, it modulated sperm metabolism and binding by regulating susceptible surface proteins. Changes in CSE1L, PFDN4, and STK39 during in vitro capacitation were confirmed using immunocytochemistry, image-based flow cytometry, and Western blotting. The results confirmed the active participation of the UPS in the extensive sperm surface proteome remodeling that occurs during boar sperm capacitation. This work will help us to identify new pharmacological mechanisms to positively or negatively modulate sperm fertilizing ability in food animals and humans. |
| first_indexed | 2024-03-11T02:41:34Z |
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| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-11T02:41:34Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
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| series | Biomolecules |
| spelling | doaj.art-2f8e3737de7946a3b94b9f78f2a73d0a2023-11-18T09:31:42ZengMDPI AGBiomolecules2218-273X2023-06-0113699610.3390/biom13060996The Ubiquitin-Proteasome System Participates in Sperm Surface Subproteome Remodeling during Boar Sperm CapacitationMichal Zigo0Karl Kerns1Peter Sutovsky2Division of Animal Science, University of Missouri, Columbia, MO 65211, USADivision of Animal Science, University of Missouri, Columbia, MO 65211, USADepartment of Animal Science, Iowa State University, Ames, IA 50011, USASperm capacitation is a complex process endowing biological and biochemical changes to a spermatozoon for a successful encounter with an oocyte. The present study focused on the role of the ubiquitin–proteasome system (UPS) in the remodeling of the sperm surface subproteome. The sperm surface subproteome from non-capacitated and in vitro capacitated (IVC) porcine spermatozoa, with and without proteasomal inhibition, was selectively isolated. The purified sperm surface subproteome was analyzed using high-resolution, quantitative liquid chromatography–mass spectrometry (LC-MS) in four replicates. We identified 1680 HUGO annotated proteins, out of which we found 91 to be at least 1.5× less abundant (<i>p</i> < 0.05) and 141 to be at least 1.5× more abundant (<i>p</i> < 0.05) on the surface of IVC spermatozoa. These proteins were associated with sperm capacitation, hyperactivation, metabolism, acrosomal exocytosis, and fertilization. Abundances of 14 proteins were found to be significantly different (<i>p</i> < 0.05), exceeding a 1.5-fold abundance between the proteasomally inhibited (100 µM MG132) and vehicle control (0.2% ethanol) groups. The proteins NIF3L1, CSE1L, NDUFB7, PGLS, PPP4C, STK39, and TPRG1L were found to be more abundant; while BPHL, GSN, GSPT1, PFDN4, STYXL1, TIMM10, and UBXN4 were found to be less abundant in proteasomally inhibited IVC spermatozoa. Despite the UPS having a narrow range of targets, it modulated sperm metabolism and binding by regulating susceptible surface proteins. Changes in CSE1L, PFDN4, and STK39 during in vitro capacitation were confirmed using immunocytochemistry, image-based flow cytometry, and Western blotting. The results confirmed the active participation of the UPS in the extensive sperm surface proteome remodeling that occurs during boar sperm capacitation. This work will help us to identify new pharmacological mechanisms to positively or negatively modulate sperm fertilizing ability in food animals and humans.https://www.mdpi.com/2218-273X/13/6/996ubiquitin–proteasome systemspermatozoonsperm capacitationsperm surface proteinCSE1LPFDN4 |
| spellingShingle | Michal Zigo Karl Kerns Peter Sutovsky The Ubiquitin-Proteasome System Participates in Sperm Surface Subproteome Remodeling during Boar Sperm Capacitation Biomolecules ubiquitin–proteasome system spermatozoon sperm capacitation sperm surface protein CSE1L PFDN4 |
| title | The Ubiquitin-Proteasome System Participates in Sperm Surface Subproteome Remodeling during Boar Sperm Capacitation |
| title_full | The Ubiquitin-Proteasome System Participates in Sperm Surface Subproteome Remodeling during Boar Sperm Capacitation |
| title_fullStr | The Ubiquitin-Proteasome System Participates in Sperm Surface Subproteome Remodeling during Boar Sperm Capacitation |
| title_full_unstemmed | The Ubiquitin-Proteasome System Participates in Sperm Surface Subproteome Remodeling during Boar Sperm Capacitation |
| title_short | The Ubiquitin-Proteasome System Participates in Sperm Surface Subproteome Remodeling during Boar Sperm Capacitation |
| title_sort | ubiquitin proteasome system participates in sperm surface subproteome remodeling during boar sperm capacitation |
| topic | ubiquitin–proteasome system spermatozoon sperm capacitation sperm surface protein CSE1L PFDN4 |
| url | https://www.mdpi.com/2218-273X/13/6/996 |
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