To explore the isomerization levels of proteins in the seed, we performed a mass spectrometry-based proteomics analysis of the seed samples from the Nip and Daat1 groups. We achieved a robust workflow with high proteome depth, quantifying 1925 nip and 1868 Daat1 proteins, of which 1702 were shared proteins. By screening the proteins with the algorithm, both the isomerization levels and degrees of proteins were altered in daat1 compared with wild type.