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Disruption of Protein Phosphatase 1 complexes using bioportides as a novel approach to target sperm motility
Vieira Silva, Joana João Freitas, Maria Santiago, Joana Jones, Sarah Guimarães, Sofia Vijayaraghavan, Srinivasan Publicover, Steven Colombo, Giorgio Howl, John Fardilha, Margarida
Vieira Silva, Joana
João Freitas, Maria
Santiago, Joana
Jones, Sarah
Guimarães, Sofia
Vijayaraghavan, Srinivasan
Publicover, Steven
Colombo, Giorgio
Howl, John
Fardilha, Margarida
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2020-09-23
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Abstract
To design protein phosphatase 1 (PP1)-disrupting peptides covalently coupled to inert cell penetrating peptides (CPPs) as sychnologically-organized bioportide constructs as a strategy to modulate sperm motility. Design: Experimental study. Setting: Academic research laboratory. Patients/Animals: Normozoospermic men providing samples for routine analysis and Holstein Frisian bulls. Intervention(s): None. Main Outcome Measure(s): Effect of the bioportides on the activity and interactions of PP1γ2 – a PP1 isoform expressed exclusively in testicular germ cells and sperm - and on sperm vitality and motility. Results: PP1‐disrupting peptides were designed based on the sequences from (i) a sperm-specific PP1 interactor (A kinase anchor protein 4, AKAP4) and (ii) a PP1 inhibitor (protein phosphatase inhibitor 2, PPP1R2). Those sequences were covalently coupled to inert CPPs as bioportide constructs, which were successfully delivered to the flagellum of sperm cells to induce a marked impact upon PP1γ2 activity and sperm motility. Molecular modelling studies further facilitated the identification of an optimized PP1 binding sequence and enabled the development of a Modified Stop Sperm (MSS1) bioportide with reduced size and increased potency of action. Additionally, a bioportide mimetic of the unique 22-amino acid C-terminus of PP1γ2 accumulated within spermatozoa to significantly reduce sperm motility and further define the PP1γ2-specific interactome. Conclusion: These investigations demonstrate the utility of CPPs to deliver peptide sequences 53 that target unique protein-protein interactions in spermatozoa to achieve a significant impact upon 54 spermatozoa motility, a key prognostic indicator of male fertility.
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Vieira Silva, J., João Freitas, M., Santiago, J., Jones, S. et al. (2021) Disruption of Protein Phosphatase 1 complexes using bioportides as a novel approach to target sperm motility, Fertility and Sterility, 115 (2), pp. 348-362. https://doi.org/10.1016/j.fertnstert.2020.08.013
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Journal article
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en
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This is an accepted manuscript of an article published by Elsevier in Fertility and Sterility on 23/09/2020, available online: https://doi.org/10.1016/j.fertnstert.2020.08.013
The accepted version of the publication may differ from the final published version.
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0015-0282
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This work was supported by FEDER funds through the “Programa Operacional Competitividade e Internacionalização COMPETE 2020” and by National Funds through the FCT Fundação para a Ciência e Tecnologia (PTDB/BBB-BQB/3804/2014). We are thankful to iBiMED (UIDB/04501/2020 and POCI-01-0145-FEDER-007628) for supporting this project. This work was also supported by an individual grant from FCT to J.V.S. (SFRH/BPD/123155/2016).