Dataset Information

Interactome analysis of Dap20-linked diUb

ABSTRACT: Biotinylated diUb species and monoubiquitin were immobilized on streptavidin-coated magnetic beads and subjected to an in vitro pulldown assay using HCT116 whole-cell extracts.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Takuya Tomita

LAB HEAD: Yasushi Saeki

PROVIDER: PXD069069 | Pride | 2025-12-15

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20250422_Tomita_Zoom_40SPD.msf	Msf
	20250422_Tomita_Zoom_40SPD.pep.xml	Pepxml
	20250422_Tomita_Zoom_40SPD_A01.raw	Raw
	20250422_Tomita_Zoom_40SPD_A02.raw	Raw
	20250422_Tomita_Zoom_40SPD_A03.raw	Raw

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Project description:Predicting male fertility remains a major challenge in livestock industry. Seminal plasma contains a heterogeneous population of extracellular vesicles (sEVs) that are involved in the regulation of sperm functionality and may therefore be involved in male fertility. The aim of this study was to evaluate whether the protein load of sEVs differs between boars showing differences in fertility when used for artificial insemination (AI). An in-depth quantitative proteomic analysis was performed on two subsets of sEVs of different size (small [S] and large [L]) isolated from boars with different reproductive performance as assessed by farrowing rate (FR) and litter size (LS). Seminal plasma samples from 18 AI-boars (three per boar) from four fertility groups: high FR (H-FR, n=5), low FR (L-FR, n=5), large LS (L-LS, n=4), and small LS (S-LS, n=4) were used. Seminal plasma samples from each fertility group were randomly mixed resulting in three seminal plasma pools per fertility group. Seminal EVs were isolated by size exclusion chromatography and characterized according to MISEV2023 guidelines. Proteomic analysis was performed using a Bruker timsTOF fleX™ instrument with dia PASEF technology. A total of 470 and 726 proteins were quantified in S-sEVs, and 1,801 and 1,834 proteins in L-sEVs from boars with differences in FR and LS, respectively. Differential protein abundance analysis revealed three and 12 proteins quantified (P ≤ 0.05) only in S-sEVs and L-sEVs from L-FR boar samples, and 37 and five proteins quantified (P ≤ 0.05) only in S-sEVs and L-sEVs from L-LS boar samples, respectively. For proteins quantified in all fertility groups, the comparison between L-FR and H-FR boar samples revealed 4 and 40 differentially abundant proteins (DAPs; log2 fold change ≥ ±1, P ≤ 0.05) in S-sEVs and L-sEVs, respectively. When comparing L-LS and S-LS boar samples, 10 and 47 DAPs were identified in S-sEVs and L-sEVs, respectively. These results show that the protein load of sEVs varies between more and less fertile boars and that the DAPs are different between S-sEVs and L-sEVs. These results would support that sEVs proteins would be involved in the fertility of boars and that those that are exclusively and/or differentially abundant between more and less fertile boars could be candidates for fertility biomarkers

Dataset Information

Interactome analysis of Dap20-linked diUb

Dataset's files

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