Sh3rf3 is an ASD-associated gene,Sh3rf3 deficiency reduces excitatory synaptic strength in prefrontal cortex and decreases the readily releasable vesicle pool and attenuated vesicle replenishment.To delve into the underlying mechanisms for impaired excitatory transmission, we performed immunoprecipitation mass spectrometry (IP-MS) in Sh3rf3-3×flag knock-in mice we generated. Gene Ontology analysis of proteins that interacted with SH3RF3 indicated that they were highly associated with synaptic transmission and vesicle recycling.