1RDQ

Hydrolysis of ATP in the crystal of Y204A mutant of cAMP-dependent protein kinase

1RDQ の概要

• エントリーDOI: 10.2210/pdb1rdq/pdb
• 関連するPDBエントリー: 1ATP
• 分子名称: cAMP-dependent protein kinase, alpha-catalytic subunit, cAMP-dependent protein kinase inhibitor, alpha form, PHOSPHATE ION, ... (9 entities in total)
• 機能のキーワード: camp-dependent protein kinase, catalytic mechanism, atp hydrolysis, two nucleotide states, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• 由来する生物種: Mus musculus (house mouse); 詳細
• 細胞内の位置: Cytoplasm . Isoform 2: Cell projection, cilium, flagellum : P05132
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 44079.86

構造登録者

Yang, J.,Ten Eyck, L.F.,Xuong, N.H.,Taylor, S.S. (登録日: 2003-11-05, 公開日: 2004-04-13, 最終更新日: 2024-10-30)

主引用文献

Yang, J.,Ten Eyck, L.F.,Xuong, N.H.,Taylor, S.S.
Crystal Structure of a cAMP-dependent Protein Kinase Mutant at 1.26A: New Insights into the Catalytic Mechanism.
J.Mol.Biol., 336:473-487, 2004

PubMed Abstract: The catalytic subunit of cAMP-dependent protein kinase has served as a paradigm for the entire kinase family. In the course of studying the structure-function relationship of the P+1 loop (Leu198-Leu205) of the kinase, we have solved the crystal structure of the Tyr204 to Ala mutant in complexes with Mg.ATP and an inhibitory peptide at 1.26A, with overall structure very similar to that of the wild-type protein. However, at the nucleotide binding site, ATP was found largely hydrolyzed, with the products ADP-PO(4) retained in the structure. High-resolution refinement suggests that 26% of the molecules contain the intact ATP, whereas 74% have the hydrolyzed products. The observation of the substrate and product states in the same structure adds significant information to our understanding of the phosphoryl transfer process. Structural examination of the mutation site substantiates and extends the emerging concept that the hydrophobic core in the large lobe of the kinase might serve as a stable platform for anchoring key segments involved in catalysis. We propose that Tyr204 is critical for anchoring the P+1 loop to the core. Further analysis has highlighted two major connections between the P+1 loop and the catalytic loop (Arg165-Asn171). One emphasizes the hydrophobic packing of Tyr204 and Leu167 mediated through residues from the alphaF-helix, recently recognized as a signal integration motif, which together with the alphaE-helix forms the center of the hydrophobic core network. The other connection is mediated by the hydrogen bond interaction between Thr201 and Asp166, in a substrate-dependent manner. We speculate that the latter interaction may be important for the kinase to sense the presence of substrate and prepare itself for the catalytic reaction. Thus, the P+1 loop is not merely involved in substrate binding; it mediates the communication between substrate and catalytic residues.

PubMed: 14757059
DOI: 10.1016/j.jmb.2003.11.044

実験手法

X-RAY DIFFRACTION (1.26 Å)