Abstract
We have overexpressed an 8.5-kDa mouse Ca(2+)/calmodulin kinase II inhibitor alpha protein (mCaMKIINalpha) in Escherichia coli and demonstrate that the recombinant protein is a potent inhibitor of Ca(2+)/calmodulin kinase II (CaMKII) in vitro. However, antibodies raised against recombinant mCaMKIINalpha react with an approximately 37-kDa protein present in mouse brain. The pattern of expression of the approximately 37-kDa protein is similar to that of mCaMKIINalpha mRNA as both are expressed in normal but not Japanese encephalitis virus (JEV)-infected mouse brain. Subcellular localization studies indicate that the approximately 37-kDa protein is present in the post-synaptic density (PSD) where mCaMKIIalpha is known to perform key regulatory functions. We conclude that the approximately 37-kDa protein identified in this study is mCaMKIINalpha and its localization in the PSD indicates a novel role for this protein in the regulation of neuronal CaMKIIalpha.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Brain / enzymology*
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Brain Chemistry / physiology*
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Calcium Signaling / physiology
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Calcium-Calmodulin-Dependent Protein Kinase Type 2
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Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
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Carrier Proteins / genetics
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Carrier Proteins / metabolism*
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Encephalitis, Japanese / enzymology
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Encephalitis, Japanese / genetics
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Intracellular Signaling Peptides and Proteins
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Mice
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Molecular Sequence Data
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Neurons / enzymology*
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Neurons / ultrastructure
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RNA, Messenger / metabolism
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Synaptic Membranes / enzymology
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Synaptic Membranes / genetics
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Synaptic Membranes / ultrastructure
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Synaptic Transmission / physiology
Substances
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Camk2n2 protein, mouse
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Carrier Proteins
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Intracellular Signaling Peptides and Proteins
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RNA, Messenger
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Recombinant Fusion Proteins
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Calcium-Calmodulin-Dependent Protein Kinase Type 2
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Calcium-Calmodulin-Dependent Protein Kinases