Identification of a Potential Tyrosine Phosphorylation Site on the NR2B Subunit of the N-methyl-D-aspartate Receptor

NMDA 수용체 아단위 2B의 Tyrosine 인산화 위치의 동정

  • Il Soo Moon (Department of Anatomy, College of Medicine, Dongguk University) ;
  • Yong Wook Jung (Department of Anatomy, College of Medicine, Dongguk University) ;
  • Bok Hyun Ko (Department of Anatomy, College of Medicine, Dongguk University)
  • Published : 1998.12.01

Abstract

The 2B subunit of N-methyl-D-aspartate (NMDA) receptors (NR2B) is the major phosphotyrosine-containing pro-tein in the postsynaptic density (PSD). In order to identify the site for tyrosine phosphorylation on NR2B, a mass spectrometry was applied on tryptic and endolys-C peptides. The NR2B subunit was isolated from N-octyl glucoside (NOG)-insoluble PSD fraction through SDS-PAGE and electroelution. The eluted protein was confirmed to be NR2B and phosphorylated on tyrosine by its cognate antibody and phosphotyrosine-specific antibody. By matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry of the peptides generated by digesting the eluted NR2B with trysin or endolys-C, a potential site for tyrosine phosphorylation could be identified as Tyr-1304.