title: Structural Basis of Cyclic Nucleotide Selectivity in cGMP-dependent Protein Kinase II.
abstract: Membrane-bound cGMP-dependent protein kinase (PKG) II is a key regulator of bone growth, renin secretion, and memory formation. Despite its crucial physiological roles, little is known about its cyclic nucleotide selectivity mechanism due to a lack of structural information. Here, we find that the C-terminal cyclic nucleotide binding (CNB-B) domain of PKG II binds cGMP with higher affinity and selectivity when compared with its N-terminal CNB (CNB-A) domain. To understand the structural basis of cGMP selectivity, we solved co-crystal structures of the CNB domains with cyclic nucleotides. Our structures combined with mutagenesis demonstrate that the guanine-specific contacts at Asp-412 and Arg-415 of the αC-helix of CNB-B are crucial for cGMP selectivity and activation of PKG II. Structural comparison with the cGMP selective CNB domains of human PKG I and Plasmodium falciparum PKG (PfPKG) shows different contacts with the guanine moiety, revealing a unique cGMP selectivity mechanism for PKG II.
authors.affiliation: From the Structural and Computational Biology and Molecular Biophysics Program.
authors.forename: James C
authors.initials: JC
authors.lastname: Campbell
authors.affiliation: Department of Pharmacology, and; the Department of Biochemistry, University of Kassel, Kassel, Hesse 34132, Germany.
authors.forename: Jeong Joo
authors.initials: JJ
authors.lastname: Kim
authors.affiliation: the Department of Biochemistry & Cell Biology, Rice University, Houston, Texas 77005.
authors.forename: Kevin Y
authors.initials: KY
authors.lastname: Li
authors.affiliation: Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030.
authors.forename: Gilbert Y
authors.initials: GY
authors.lastname: Huang
authors.affiliation: Department of Pharmacology, and.
authors.forename: Albert S
authors.initials: AS
authors.lastname: Reger
authors.affiliation: the Department of Biological Science and Technology, the University of Tokushima Graduate School, Tokushima 770-8506, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Matsuda
authors.affiliation: the Berkeley Center for Structural Biology, Lawrence Berkeley National Laboratory, Berkeley, California 94720.
authors.forename: Banumathi
authors.initials: B
authors.lastname: Sankaran
authors.affiliation: the Department of Biochemistry and Molecular Biology, The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030, and.
authors.forename: Todd M
authors.initials: TM
authors.lastname: Link
authors.affiliation: the Department of Biological Science and Technology, the University of Tokushima Graduate School, Tokushima 770-8506, Japan.
authors.forename: Keizo
authors.initials: K
authors.lastname: Yuasa
authors.affiliation: From the Structural and Computational Biology and Molecular Biophysics Program,; the Department of Biochemistry and Molecular Biology, The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030, and.
authors.forename: John E
authors.initials: JE
authors.lastname: Ladbury
authors.affiliation: the Department of Medicine, University of California, San Diego, La Jolla, California 92093.
authors.forename: Darren E
authors.initials: DE
authors.lastname: Casteel
authors.affiliation: From the Structural and Computational Biology and Molecular Biophysics Program,; Department of Pharmacology, and; Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030,. Electronic address: ckim@bcm.edu.
authors.forename: Choel
authors.initials: C
authors.lastname: Kim
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D018360:Crystallography, X-Ray; D000242:Cyclic AMP / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism*; D062967:Cyclic GMP-Dependent Protein Kinase Type II / Q000737:chemistry* / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity
chemical_list: D000242:Cyclic AMP; D062967:Cyclic GMP-Dependent Protein Kinase Type II; D006152:Cyclic GMP
keywords: NO-cGMP signaling; X-ray crystallography; allosteric regulation; cGMP-dependent protein kinase; cyclic nucleotide; cyclic nucleotide-binding domain (CNB); ligand-binding protein; protein kinase G (PKG); receptor structure-function; second messenger; serine/threonine protein kinase