Protein microarray analysis identifies cyclic nucleotide phosphodiesterase as an interactor of Nogo-A.
Nogo-A, a neurite outgrowth inhibitor, is expressed exclusively on oligodendrocytes and neurons in the CNS. The central domain of Amino-Nogo spanning amino acids 567-748 in the human Nogo-A designated NIG, mediates persistent inhibition of axonal outgrowth and induces growth cone collapse by signaling through an as yet unidentified NIG receptor. We identified 82 NIG-interacting proteins by screening a high-density human protein microarray composed of 5000 proteins with a recombinant NIG protein as a probe. Following an intensive database search, we selected 12 neuron/oligodendrocyte-associated NIG interactors. Among them, we verified the molecular interaction of NIG with 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNP), a cell type-specific marker of oligodendrocytes, by immunoprecipitation and cell imaging analysis. Although CNP located chiefly in the cytoplasm of oligodendrocytes might not serve as a cell-surface NIG receptor, it could act as a conformational stabilizer for the intrinsically unstructured large segment of Amino-Nogo.
Sumiyoshi
Kenta
K
Department of Bioinformatics and Molecular Neuropathology, Meiji Pharmaceutical University, Tokyo 204-8588, Japan.
Obayashi
Shinya
S
Tabunoki
Hiroko
H
Arima
Kunimasa
K
Satoh
Jun-ichi
J
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D030562:Databases, Protein; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008875:Middle Aged; D009185:Myelin Proteins / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D000070798:Nogo Proteins; D009836:Oligodendroglia / Q000378:metabolism; D040081:Protein Array Analysis; D011994:Recombinant Proteins / Q000378:metabolism; D015203:Reproducibility of Results
D009185:Myelin Proteins; D000070798:Nogo Proteins; C000603199:RTN4 protein, human; D011994:Recombinant Proteins; C000603201:Rtn4 protein, mouse; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases