title: High-level expression of functional cGMP-dependent protein kinase using the baculovirus system.
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 I alpha was constructed and purified. Infected insect cells synthesized large amounts of soluble and biologically active cGMP kinase I alpha 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 I alpha should provide an excellent tool to study further the structure and function of cGMP kinase.
authors.affiliation: Institut für Pharmakologie und Toxikologie der Technischen Universität München, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Feil
authors.forename: S
authors.initials: S
authors.lastname: Müller
authors.forename: F
authors.initials: F
authors.lastname: Hofmann
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D017869:Cyclic GMP-Dependent Protein Kinases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009036:Moths; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
chemical_list: D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D017869:Cyclic GMP-Dependent Protein Kinases