title: Crystal structure of the catalytic subunit of cAMP-dependent protein kinase complexed with MgATP and peptide inhibitor.
abstract: The structure of a ternary complex of the catalytic subunit of cAMP-dependent protein kinase, MgATP, and a 20-residue inhibitor peptide was determined at a resolution of 2.7 A using the difference Fourier technique starting from the model of the binary complex (Knighton et al., 1991a). The model of the ternary complex was refined using both X-PLOR and TNT to an R factor of 0.212 and 0.224, respectively. The orientation of the nucleotide and the interactions of MgATP with numerous conserved residues at the active site of the enzyme are clearly defined. The unique protein kinase nucleotide binding site consists of a five-stranded antiparallel beta-sheet with the base buried in a hydrophobic site along beta-strands 1 and 2 and fixed by hydrogen bonds to the N6 amino and N7 nitrogens. The small lobe secures the nucleotide via a glycine-rich loop and by ion pairing with Lys72 and Glu91. While the small lobe fixes the nontransferable alpha- and beta-phosphates in this inhibitor complex, the gamma-phosphate is secured by two Mg2+ ions and interacts both directly and indirectly with several residues in the large lobe--Asp184, Asn171, Lys168. Asp166 is positioned to serve as a catalytic base. The structure is correlated with previous chemical evidence, and the features that distinguish this nucleotide binding motif from other nucleotide binding proteins are delineated.
authors.affiliation: Department of Chemistry, University of California, San Diego, La Jolla 92093.
authors.forename: J
authors.initials: J
authors.lastname: Zheng
authors.forename: D R
authors.initials: DR
authors.lastname: Knighton
authors.forename: L F
authors.initials: LF
authors.lastname: ten Eyck
authors.forename: R
authors.initials: R
authors.lastname: Karlsson
authors.forename: N
authors.initials: N
authors.lastname: Xuong
authors.forename: S S
authors.initials: SS
authors.lastname: Taylor
authors.forename: J M
authors.initials: JM
authors.lastname: Sowadski
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002384:Catalysis; D003198:Computer Simulation; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D011480:Protease Inhibitors / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000737:chemistry* / Q000378:metabolism; D012266:Ribose / Q000737:chemistry; D014961:X-Ray Diffraction
chemical_list: D046911:Macromolecular Substances; D010446:Peptide Fragments; D011480:Protease Inhibitors; D047428:Protein Kinase Inhibitors; C518603:protein kinase inhibitor peptide (5-24); D012266:Ribose; D000255:Adenosine Triphosphate; D011494:Protein Kinases