Proteomic analysis

Identification of the interacting partners of Zip was done by co-immunoprecipitation of the interacting proteins from GFP-Zip over-expression protein lysate with anti-GFP antibody followed by in-solution trypsin digestion and analysis by high-resolution mass spectrometry. The peptides identified in the HRMS screening were sorted on the basis of the sequence score (> 40). STRING database (https://string-db.org/) was used to generate the protein interactome based on identification of Zip interacting partners. Functional enrichment of the biological processes in the identified network represented as log 10 having p values with Benjamini–Hochberg correction has been shown in the bar graph. High-Resolution Accurate Mass Spectrometry System (Orbitrap Eclipse Tribrid Mass Spectrometer) from Thermo Fischer Scientific was used for the MS screening of Zip interacting proteins.