Mutagenesis of Thr-286 in monomeric Ca 2 + / calmodulin-dependent protein kinase II eliminates Ca 2 + / calmodulin-independent activity ( bacterial expression / autophosphorylation / pseudosubstrate )

Abstract

We have examined the role of Thr-286 autophosphorylation in the autoregulation of Ca2+/calmodulindependent protein kinase U. Using site-directed mutagenesis, we have substituted alanine or serine for Thr-286, or isoleucine for Arg-283, in the 50-kDa subunit of the kinase and expressed each protein in bacteria. Activation and autophosphorylation of all four… (More)

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