Amp cp

Drosophila full-length CaMKII (R3 isoform;^{58 (link)}) was a gift of Leslie Griffith (Brandeis University, Massachusetts, U.S.A.) while full-length Drosophila EAG (GenBank: M61157) was a gift of Dianne Papazian (UCLA, California, U.S.A.). Nucleotides (ATP (Sigma), AMPPNP (Jena Biosciences), ADP (Sigma) and AMPCP (Sigma)) were processed according to manufactures’ instructions. Stock solutions at 50 mM of ATP, AMPPNP and ADP were freshly prepared in either 1 M Tris pH 7.5 or 1 M HEPES pH 7.5. Stock solution of AMPCP at 50 mM in 1 M HEPES pH 7.5 was stored at −20 °C. λ-phosphatase (NEB) was processed according to manufacturer’s instructions, aliquoted and stored at −80 °C. Stock solutions of reagents used in the ATPase assay were prepared as follows: phosphoenolpyruvate (PEP; Sigma) at 50 mM in 50 mM HEPES pH 7.5; nicotinamide adenine dinucleotide, reduced (NADH; Sigma) at 6 mM in 50 mM HEPES pH 7.5; peptide syntide (GenScript) was prepared in water at 2.5 mM. All these solutions were kept aliquoted at −80 °C. The pyruvate kinase/lactate dehydrogenase mix (Sigma) was at 600–1000 units ml⁻¹ and 900–1400 units ml⁻¹ activity, respectively, and kept at −20 °C.