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Using siRNA against various forms of the G protein-coupled receptor kinase (GRK) family, DAMGO was demonstrated to phosphorylate T370 and S375 by GRK2 and 3 while morphine increases S375 phosphorylation by GRK5 (Doll et al., In vivo , using KO mice for either GRK3 or 5, morphine rather promotes MOR phosphorylation at S375 by both kinases while only GRK3 was required for fentanyl-induced MOR phosphorylation (Glck et al., The tyrosine kinase Src was also shown to phosphorylate MOR at Y336, located in the NPXXY motif, after sustained morphine treatment followed by naloxone (Zhang et al., 2009)

Mass Spectrometry: Data-dependent MS/MS analysis was performed using a Q Exactive Orbitrap mass spectrometer (Thermo Scientific)