Fig. 1.

Experimental overview. (A) Fertilization and egg activation release metaphase-arrested oocytes into anaphase and initiate the slow block to polyspermy, sister chromatid separation, remodeling of the specialized egg extracellular matrix, and inflation of the perivitelline space, among other processes. We mimic fertilization and trigger egg activation via electric shock to maximize synchrony. (B) After activation, eggs were collected every 2 min, lysed, and digested with proteases. Samples were barcoded using tandem mass tags (TMTs) and multiplexed. Protein and phosphosite dynamics were measured with mass spectrometry-based proteomics. Phosphorylated peptides were enriched using TiO₂, and the column flow-through peptides were used for protein analysis. (C) Modified waterfall plots displaying the trend of every protein and phosphosite in the dataset, normalized to the first time point and ordered first by clusters (see below) and then in ascending order within each cluster. (D and E) K-means clustering to summarize dynamic classes of the protein (D) and phosphorylation time series (E) (SI Appendix, SI Methods). Bold line represents the centroid of the cluster, while the gray lines are individual time courses (normalized to the time course means).