Dataset Information

Proteomics of phosphorylation and protein dynamics during fertilization and meiotic exit in the Xenopus egg

ABSTRACT: Fertilization triggers release from meiotic arrest and initiates events that prepare for the ensuing developmental program. Protein degradation and phosphorylation are known to regulate protein activity during this process. However, the full extent of protein loss and phospho-regulation is still unknown. We examined absolute protein and phospho-site dynamics after fertilization by mass spectrometry-based proteomics. To do this, we developed a new approach for calculating the stoichiometry of phospho-sites from multiplexed proteomics compatible with dynamic, stable and multi-site phosphorylation. Overall, the data suggest that degradation is limited to a few low abundance proteins. However, this degradation in part promotes extensive dephosphorylation that occurs over a wide range of abundances during meiotic exit. We also show that eggs release a large amount of protein into the medium just after fertilization, most likely related to the blocks to polyspermy. Concomitantly, there is a substantial increase in phosphorylation likely tied to calcium activated kinases. We identify putative degradation targets as well as new components of the block to polyspermy. The analytical approaches demonstrated here are broadly applicable to studies of dynamic biological systems.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Xenopus Laevis (african Clawed Frog)

TISSUE(S): Egg, Oocyte

SUBMITTER: Marc Presler

LAB HEAD: Marc W. Kirschner

PROVIDER: PXD006639 | Pride | 2017-11-29

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	c00406_eap3r_0_scx.raw	Raw
	c00406_eap3r_0_scx_20160318_check_phospho.tgz	Other
	c00407_eap3r_5_scx.raw	Raw
	c00407_eap3r_5_scx_20160318_check_phospho.tgz	Other
	c00408_eap3r_10_scx.raw	Raw

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Proteomics of phosphorylation and protein dynamics during fertilization and meiotic exit in the <i>Xenopus</i> egg.

Presler Marc M Van Itallie Elizabeth E Klein Allon M AM Kunz Ryan R Coughlin Margaret L ML Peshkin Leonid L Gygi Steven P SP Wühr Martin M Kirschner Marc W MW

Proceedings of the National Academy of Sciences of the United States of America 20171128 50

Fertilization releases the meiotic arrest and initiates the events that prepare the egg for the ensuing developmental program. Protein degradation and phosphorylation are known to regulate protein activity during this process. However, the full extent of protein loss and phosphoregulation is still unknown. We examined absolute protein and phosphosite dynamics of the fertilization response by mass spectrometry-based proteomics in electroactivated eggs. To do this, we developed an approach for cal ...[more]

PMID: 29183978

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