A Mass Spectrometry-based Proteomic Analysis of Homer2-interacting Proteins in the Mouse Brain
Goulding, S.P.; Szumlinski, K.K.; Contet, C.; et al.
Scaffolding proteins of the Homer family are recognized as important indicators of several psychiatric disorders. To better understand their molecular function toward improved therapeutic strategies, the interactions of Homer proteins must be identified. Here, Goulding and colleagues employed co-immunoprecipitation (via rabbit polyclonal anti-Homer2 antibody) with nano-LC-MS/MS (DDA and SRM acquisition for discovery and verification, respectively) to identify interacting Homer2 proteins in mouse brain tissue (C57BL/6 strain). To help determine retention times, peptides from 15N Apolipoprotein A-I were used as iRT standards. Overall, the verification experiments revealed 18 Homer2-interacting proteins of high confidence, with 15 of those being novel interactors stemming largely from the N-methyl-D-aspartate receptor (NMDAR) signaling pathway (e.g., GluN1 and GluN2B). The results help demonstrate the functional implication of Homer2 in signal transduction.