Regulatory phosphorylation of serine-15 in maize phospho enolpyruvate carboxylase by a C 4-leaf protein-serine kinase

Abstract

We have recently reported that the light-induced changes in the enzymatic and regulatory properties of maize leaf phospho enolpyruvate carboxylase are attributed to the regulatory seryl phosphorylation of this C 4-photosynthesis enzyme. In the present study, the darkform target enzyme was phosphorylated/activated in vitro by a maize leaf protein-serine kinase, and the 32P-labeled regulatory site phosphopeptide was purified from a tryptic digest by metal-ion affinity and reversed-phase chromatography. Automated Edman degradation analysis by covalent protein sequencing technology revealed that the amino acid sequence of this phosphoseryl peptide is His-His-Ser(P)-Ile-Asp-Ala-Gln-Leu-Arg. This nonapeptide, which corresponds exactly to residues 13–21 in the deduced primary sequence of the maize leaf carboxylase, is far removed from recently identified active-site cysteine (Cys-553) and lysine (Lys-606) residues in the C-terminal region of the primary structure. Comparative analysis of the deduced N-terminal sequences of C 3-, C 4-, and Crassulacean acid metabolism (CAM)-leaf phospho enolpyruvate carboxylases suggests that the motif of Lys/Arg-X-X-Ser is an important structural requirement of the C 4- and CAM-leaf protein-serine kinases.