Analysis of phosphopeptide changes as spermatozoa acquire functional competence in the epididymis demonstrates changes in the post-translational modification of izumo1

Mark A. Baker, Louise Hetherington, Anita Weinberg, Nenad Naumovski, Tony Velkov, Matthias Pelzing, Sebastiaan Dolman, Mark R. Condina, R. John Aitken

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Spermatozoa are functionally inert when they emerge from the testes. Functional competence is conferred upon these cells during a post-testicular phase of sperm maturation in the epididymis. Remarkably, this functional transformation of epididymal spermatozoa occurs in the absence of nuclear gene transcription or protein translation. To understand the cellular mechanisms underpinning epididymal maturation, we have performed a label-free, MS-based, comparative quantification of peptides from caput, corpus and caudal epididymal spermatozoa. In total, 68 phosphopeptide changes could be detected during epididymal maturation corresponding to the identification of 22 modified proteins. Included in this list are the sodium-bicarbonate cotransporter, the sperm specific serine kinase 1, AKAP4 and protein kinase A regulatory subunit. Furthermore, four phosphopeptide changes came from Izumo1, the sperm-egg fusion protein, in the cytoplasmic segment of the protein. 2D-PAGE confirmed that Izumo1 is post-translationally modified during epididymal transit. Interestingly, phosphorylation on Izumo1 was detected on residue S339 in the caput and corpus but not caudal cells. Furthermore, Izumo1 exhibited four phosphorylated residues when spermatozoa reached the cauda, which were absent from caput cells. A model is advanced suggesting that these phospho-regulations are likely to act as a scaffold for the association of adaptor proteins with Izumo1 as these cells prepare for fertilization.

Original languageEnglish
Pages (from-to)5252-5264
Number of pages13
JournalJournal of Proteome Research
Volume11
Issue number11
DOIs
Publication statusPublished - 2 Nov 2012
Externally publishedYes

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Phosphopeptides
Epididymis
Post Translational Protein Processing
Mental Competency
Spermatozoa
Proteins
Sodium-Bicarbonate Symporters
Phosphorylation
Sperm Maturation
Protein-Serine-Threonine Kinases
Egg Proteins
Transcription
Cyclic AMP-Dependent Protein Kinases
Scaffolds
Electrophoresis, Gel, Two-Dimensional
Protein Biosynthesis
Labels
Fertilization
Fusion reactions
Genes

Cite this

Baker, Mark A. ; Hetherington, Louise ; Weinberg, Anita ; Naumovski, Nenad ; Velkov, Tony ; Pelzing, Matthias ; Dolman, Sebastiaan ; Condina, Mark R. ; Aitken, R. John. / Analysis of phosphopeptide changes as spermatozoa acquire functional competence in the epididymis demonstrates changes in the post-translational modification of izumo1. In: Journal of Proteome Research. 2012 ; Vol. 11, No. 11. pp. 5252-5264.
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abstract = "Spermatozoa are functionally inert when they emerge from the testes. Functional competence is conferred upon these cells during a post-testicular phase of sperm maturation in the epididymis. Remarkably, this functional transformation of epididymal spermatozoa occurs in the absence of nuclear gene transcription or protein translation. To understand the cellular mechanisms underpinning epididymal maturation, we have performed a label-free, MS-based, comparative quantification of peptides from caput, corpus and caudal epididymal spermatozoa. In total, 68 phosphopeptide changes could be detected during epididymal maturation corresponding to the identification of 22 modified proteins. Included in this list are the sodium-bicarbonate cotransporter, the sperm specific serine kinase 1, AKAP4 and protein kinase A regulatory subunit. Furthermore, four phosphopeptide changes came from Izumo1, the sperm-egg fusion protein, in the cytoplasmic segment of the protein. 2D-PAGE confirmed that Izumo1 is post-translationally modified during epididymal transit. Interestingly, phosphorylation on Izumo1 was detected on residue S339 in the caput and corpus but not caudal cells. Furthermore, Izumo1 exhibited four phosphorylated residues when spermatozoa reached the cauda, which were absent from caput cells. A model is advanced suggesting that these phospho-regulations are likely to act as a scaffold for the association of adaptor proteins with Izumo1 as these cells prepare for fertilization.",
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Analysis of phosphopeptide changes as spermatozoa acquire functional competence in the epididymis demonstrates changes in the post-translational modification of izumo1. / Baker, Mark A.; Hetherington, Louise; Weinberg, Anita; Naumovski, Nenad; Velkov, Tony; Pelzing, Matthias; Dolman, Sebastiaan; Condina, Mark R.; Aitken, R. John.

In: Journal of Proteome Research, Vol. 11, No. 11, 02.11.2012, p. 5252-5264.

Research output: Contribution to journalArticle

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T1 - Analysis of phosphopeptide changes as spermatozoa acquire functional competence in the epididymis demonstrates changes in the post-translational modification of izumo1

AU - Baker, Mark A.

AU - Hetherington, Louise

AU - Weinberg, Anita

AU - Naumovski, Nenad

AU - Velkov, Tony

AU - Pelzing, Matthias

AU - Dolman, Sebastiaan

AU - Condina, Mark R.

AU - Aitken, R. John

PY - 2012/11/2

Y1 - 2012/11/2

N2 - Spermatozoa are functionally inert when they emerge from the testes. Functional competence is conferred upon these cells during a post-testicular phase of sperm maturation in the epididymis. Remarkably, this functional transformation of epididymal spermatozoa occurs in the absence of nuclear gene transcription or protein translation. To understand the cellular mechanisms underpinning epididymal maturation, we have performed a label-free, MS-based, comparative quantification of peptides from caput, corpus and caudal epididymal spermatozoa. In total, 68 phosphopeptide changes could be detected during epididymal maturation corresponding to the identification of 22 modified proteins. Included in this list are the sodium-bicarbonate cotransporter, the sperm specific serine kinase 1, AKAP4 and protein kinase A regulatory subunit. Furthermore, four phosphopeptide changes came from Izumo1, the sperm-egg fusion protein, in the cytoplasmic segment of the protein. 2D-PAGE confirmed that Izumo1 is post-translationally modified during epididymal transit. Interestingly, phosphorylation on Izumo1 was detected on residue S339 in the caput and corpus but not caudal cells. Furthermore, Izumo1 exhibited four phosphorylated residues when spermatozoa reached the cauda, which were absent from caput cells. A model is advanced suggesting that these phospho-regulations are likely to act as a scaffold for the association of adaptor proteins with Izumo1 as these cells prepare for fertilization.

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KW - titanium dioxide

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