Human spermatozoa contain multiple targets for protein S-nitrosylation

An alternative mechanism of the modulation of sperm function by nitric oxide?

Linda Lefièvre, Yongjian Chen, Sarah J. Conner, Joanna L. Scott, Steve J. Publicover, W. Christopher L. Ford, Christopher L. R. Barratt (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    105 Citations (Scopus)

    Abstract

    Nitric oxide (NO) enhances human sperm motility and capacitation associated with increased protein phosphorylation. NO activates soluble guanylyl cyclase, but can also modify protein function covalently via S-nitrosylation of cysteine. Remarkably, this mechanism remains unexplored in sperm although they depend on post-translational protein modification to achieve changes in function required for fertilisation. Our objective was to identify targets for S-nitrosylation in human sperm. Spermatozoa were incubated with NO donors and S-nitrosylated proteins were identified using the biotin switch assay and a proteomic approach using MS/MS. 240 S-nitrosylated proteins were detected in sperm incubated with S-nitroso-glutathione. Minimal levels were observed in glutathione or untreated samples. Proteins identified consistently based on multiple peptides included established targets for S-nitrosylation in other cells e.g. tubulin, GST and HSPs but also novel targets including A-kinase anchoring protein (AKAP) types 3 and 4, voltage-dependent anion-selective channel protein 3 and semenogelin 1 and 2. In situ localisation revealed S-nitrosylated targets on the postacrosomal region of the head and throughout the flagellum. Potential targets for S-nitrosylation in human sperm include physiologically significant proteins not previously reported in other cells. Their identification will provide novel insight into the mechanism of action of NO in spermatozoa.

    Original languageEnglish
    Pages (from-to)3066-3084
    Number of pages19
    JournalProteomics
    Volume7
    Issue number17
    DOIs
    Publication statusPublished - Sep 2007

    Fingerprint

    Protein S
    Spermatozoa
    Nitric Oxide
    Modulation
    Proteins
    Glutathione
    Voltage-Dependent Anion Channels
    Sperm Capacitation
    Nitric Oxide Donors
    Flagella
    Sperm Motility
    Post Translational Protein Processing
    Tubulin
    Phosphorylation
    Biotin
    Guanylate Cyclase
    Fertilization
    Proteomics
    Protein Kinases
    Cysteine

    Keywords

    • Adaptor Proteins, Signal Transducing/metabolism
    • Cysteine/metabolism
    • Humans
    • Male
    • Mitochondrial Membrane Transport Proteins
    • Nitric Oxide/metabolism
    • Nitric Oxide Donors/metabolism
    • Nitric Oxide Synthase Type III/metabolism
    • Nitroso Compounds/metabolism
    • Proteome/chemistry
    • Proteomics/methods
    • S-Nitrosoglutathione/metabolism
    • Seminal Vesicle Secretory Proteins/metabolism
    • Spermatozoa/metabolism
    • Tandem Mass Spectrometry
    • Voltage-Dependent Anion Channels/metabolism

    Cite this

    Lefièvre, Linda ; Chen, Yongjian ; Conner, Sarah J. ; Scott, Joanna L. ; Publicover, Steve J. ; Ford, W. Christopher L. ; Barratt, Christopher L. R. / Human spermatozoa contain multiple targets for protein S-nitrosylation : An alternative mechanism of the modulation of sperm function by nitric oxide?. In: Proteomics. 2007 ; Vol. 7, No. 17. pp. 3066-3084.
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    abstract = "Nitric oxide (NO) enhances human sperm motility and capacitation associated with increased protein phosphorylation. NO activates soluble guanylyl cyclase, but can also modify protein function covalently via S-nitrosylation of cysteine. Remarkably, this mechanism remains unexplored in sperm although they depend on post-translational protein modification to achieve changes in function required for fertilisation. Our objective was to identify targets for S-nitrosylation in human sperm. Spermatozoa were incubated with NO donors and S-nitrosylated proteins were identified using the biotin switch assay and a proteomic approach using MS/MS. 240 S-nitrosylated proteins were detected in sperm incubated with S-nitroso-glutathione. Minimal levels were observed in glutathione or untreated samples. Proteins identified consistently based on multiple peptides included established targets for S-nitrosylation in other cells e.g. tubulin, GST and HSPs but also novel targets including A-kinase anchoring protein (AKAP) types 3 and 4, voltage-dependent anion-selective channel protein 3 and semenogelin 1 and 2. In situ localisation revealed S-nitrosylated targets on the postacrosomal region of the head and throughout the flagellum. Potential targets for S-nitrosylation in human sperm include physiologically significant proteins not previously reported in other cells. Their identification will provide novel insight into the mechanism of action of NO in spermatozoa.",
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    author = "Linda Lefi{\`e}vre and Yongjian Chen and Conner, {Sarah J.} and Scott, {Joanna L.} and Publicover, {Steve J.} and Ford, {W. Christopher L.} and Barratt, {Christopher L. R.}",
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    Human spermatozoa contain multiple targets for protein S-nitrosylation : An alternative mechanism of the modulation of sperm function by nitric oxide? / Lefièvre, Linda; Chen, Yongjian; Conner, Sarah J.; Scott, Joanna L.; Publicover, Steve J.; Ford, W. Christopher L.; Barratt, Christopher L. R. (Lead / Corresponding author).

    In: Proteomics, Vol. 7, No. 17, 09.2007, p. 3066-3084.

    Research output: Contribution to journalArticle

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    AU - Lefièvre, Linda

    AU - Chen, Yongjian

    AU - Conner, Sarah J.

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    AU - Barratt, Christopher L. R.

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