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Monoclonal Antibody NM2 Recognizes the Protein Kinase C Phosphorylation Site in B‐50 (GAP‐43) and in Neurogranin (BICKS)
Author(s) -
Oestreicher A. Beate,
Hens Jacques J. H.,
Marquart Arnoud,
Mercken Marc,
Graan Pierre N. E.,
Zwiers Henk,
Gispen Willem Hendrik
Publication year - 1994
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1994.62030881.x
Subject(s) - neurogranin , protein kinase c , phosphorylation , peptide , monoclonal antibody , biochemistry , epitope , serine , biology , chemistry , microbiology and biotechnology , antibody , immunology
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.