Exchange protein directly activated by cAMP (EPAC) interacts with the light chain (LC) 2 of MAP1A
作者： Maria M. MAGIERA , Mona GUPTA , Catherine J. RUNDELL , Nilima SATISH , Isabelle ERNENS
DOI: 10.1042/BJ20040122
关键词: Guanine nucleotide exchange factor 、 DEP domain 、 HEK 293 cells 、 Immunoprecipitation 、 Cell culture 、 Biology 、 Transfection 、 Pleckstrin homology domain 、 Molecular biology 、 Microtubule-associated protein
摘要: Using EPAC1 (exchange protein directly activated by cAMP 1) as bait in two-hybrid screens of foetal and adult human brain libraries, we identified the LC2 (light chain 2) MAP1A (microtubule-associated 1A) a capable interaction with EPAC1. We applied an immunoprecipitation assay to demonstrate between co-transfected embryonic kidney 293 cells. EPAC2 also co-immunoprecipitated from extracts rat cerebellum. Immunolocalization cells revealed that co-localizes throughout cell body. found endogenous is immunolocalized PC12 transfected COS1 showed associated perinuclear region surrounding nucleus filamentous structures cell. Removal cAMP-binding domain (ΔcAMP-EPAC1) appeared disrupt targeting resulting more dispersed staining pattern. assay, tested ability interact ΔcAMP-EPAC1 ΔDEP-EPAC1, which lacks DEP (dishevelled, Egl-10 pleckstrin homology domain). deletion inhibited but removal had little effect. was glutathione-S-transferase-fusion pull-down not DEP, REM (Ras exchange motif) or CAT (catalytic) domains. Together our results, this suggests mediates LC2.
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