High‐level expression of functional cGMP‐dependent protein kinase using the baculovirus system

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Abstract
The understanding of the structure and function of cGMP‐dependent protein kinase (cGMP kinase) has been hindered by the difficulty to obtain large quantities of functional enzyme. A recombinant baculovirus encoding bovine cGMP kinase la was constructed and purified. Infected insect cells synthesized large amounts of soluble and biologically active cGMP kinase Iα representing up to 10% of the total cell extract protein. The recombinant enzyme had an identical apparent molecular mass, cGMP affinity and kinase activity as the native bovine lung enzyme. The high‐level expression of functional cGMP kinase Iα should provide an excellent tool to study further the structure and function of cGMP kinase.