Monoclonal Antibody NM2 Recognizes the Protein Kinase C Phosphorylation Site in B-50 (GAP-43) and in Neurogranin (BICKS)
| Autor: | Arnoud Marquart, Jacques J. H. Hens, Willem Hendrik Gispen, Henk Zwiers, Pierre N. E. De Graan, A. Beate Oestreicher, Marc Mercken |
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| Rok vydání: | 2008 |
| Předmět: |
Calmodulin
medicine.drug_class Nerve Tissue Proteins Peptide Monoclonal antibody Biochemistry Epitope Cellular and Molecular Neuroscience GAP-43 Protein medicine Animals Neurogranin Phosphorylation Rats Wistar Protein Kinase C Protein kinase C chemistry.chemical_classification Membrane Glycoproteins biology Cell Membrane Antibodies Monoclonal Molecular biology Peptide Fragments Rats Epitope mapping chemistry biology.protein Calmodulin-Binding Proteins Peptide Hydrolases Synaptosomes |
| Zdroj: | Journal of Neurochemistry. 62:881-889 |
| ISSN: | 1471-4159 0022-3042 |
| DOI: | 10.1046/j.1471-4159.1994.62030881.x |
| Popis: | Mouse monoclonal B-50 antibodies (Mabs) were screened to select a Mab that may interfere with suggested functions of B-50 (GAP-43), such as involvement in neurotransmitter release. Because the Mab NM2 reacted with peptide fragments of rat B-50 containing the unique protein kinase C (PKC) phosphorylation site at serine-41, it was selected and characterized in comparison with another Mab NM6 unreactive with these fragments. NM2, but not NM6, recognized neurogranin (BICKS), another PKC substrate, containing a homologous sequence to rat B-50 (34-52). To narrow down the epitope domain synthetic B-50 peptides were tested in ELISAs. In contrast to NM6, NM2 immunoreacted with B-50 (39-51) peptide, but not with B-50 (43-51) peptide or a C-terminal B-50 peptide. Preabsorption by B-50 (39-51) peptide of NM2 inhibited the binding of NM2 to rat B-50 in contrast to NM6. NM2 selectively inhibited phosphorylation of B-50 during endogenous phosphorylation of synaptosomal plasma membrane proteins. Preabsorption of NM2 by B-50 (39-51) peptide abolished this inhibition. In conclusion, NM2 recognizes the QASFR peptide in B-50 and neurogranin. Therefore, NM2 may be a useful tool in physiological studies of the role of PKC-mediated phosphorylation and calmodulin binding of B-50 and neurogranin. |
| Databáze: | OpenAIRE |
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