4DH1

Low temperature X-ray structure of cAMP dependent Protein Kinase A catalytic subunit with low Mg2+, ATP and IP20

4DH1 の概要

• エントリーDOI: 10.2210/pdb4dh1/pdb
• 関連するPDBエントリー: 4DH3 4DH5 4DH7 4DH8
• 分子名称: cAMP-dependent protein kinase catalytic subunit alpha, cAMP-dependent protein kinase inhibitor alpha, MAGNESIUM ION, ... (5 entities in total)
• 機能のキーワード: protein kinase, phosphorylation, peptidic inhibitor, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• 由来する生物種: Mus musculus (mouse); 詳細
• 細胞内の位置: Cytoplasm: P05132
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 43495.19

構造登録者

Kovalevsky, A.Y.,Langan, P. (登録日: 2012-01-27, 公開日: 2012-06-27, 最終更新日: 2013-03-13)

主引用文献

Kovalevsky, A.Y.,Johnson, H.,Hanson, B.L.,Waltman, M.J.,Fisher, S.Z.,Taylor, S.,Langan, P.
Low- and room-temperature X-ray structures of protein kinase A ternary complexes shed new light on its activity.
Acta Crystallogr.,Sect.D, 68:854-860, 2012

PubMed Abstract: Post-translational protein phosphorylation by protein kinase A (PKA) is a ubiquitous signalling mechanism which regulates many cellular processes. A low-temperature X-ray structure of the ternary complex of the PKA catalytic subunit (PKAc) with ATP and a 20-residue peptidic inhibitor (IP20) at the physiological Mg(2+) concentration of ∼0.5 mM (LT PKA-MgATP-IP20) revealed a single metal ion in the active site. The lack of a second metal in LT PKA-MgATP-IP20 renders the β- and γ-phosphoryl groups of ATP very flexible, with high thermal B factors. Thus, the second metal is crucial for tight positioning of the terminal phosphoryl group for transfer to a substrate, as demonstrated by comparison of the former structure with that of the LT PKA-Mg(2)ATP-IP20 complex obtained at high Mg(2+) concentration. In addition to its kinase activity, PKAc is also able to slowly catalyze the hydrolysis of ATP using a water molecule as a substrate. It was found that ATP can be readily and completely hydrolyzed to ADP and a free phosphate ion in the crystals of the ternary complex PKA-Mg(2)ATP-IP20 by X-ray irradiation at room temperature. The cleavage of ATP may be aided by X-ray-generated free hydroxyl radicals, a very reactive chemical species, which move rapidly through the crystal at room temperature. The phosphate anion is clearly visible in the electron-density maps; it remains in the active site but slides about 2 Å from its position in ATP towards Ala21 of IP20, which mimics the phosphorylation site. The phosphate thus pushes the peptidic inhibitor away from the product ADP, while resulting in dramatic conformational changes of the terminal residues 24 and 25 of IP20. X-ray structures of PKAc in complex with the nonhydrolysable ATP analogue AMP-PNP at both room and low temperature demonstrated no temperature effects on the conformation and position of IP20.

PubMed: 22751671
DOI: 10.1107/S0907444912014886

実験手法

X-RAY DIFFRACTION (2 Å)