2GNG

Protein kinase A fivefold mutant model of Rho-kinase

2GNG の概要

• エントリーDOI: 10.2210/pdb2gng/pdb
• 関連するPDBエントリー: 1Q8T 1Q8U 1Q8W 2GNF 2GNH 2GNI 2GNJ 2GNL
• 分子名称: cAMP-dependent protein kinase, alpha-catalytic subunit, cAMP-dependent protein kinase inhibitor alpha (3 entities in total)
• 機能のキーワード: pka, mutant, rho-kinase, surrogate, inhibitor, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• 由来する生物種: Bos taurus (cattle); 詳細
• 細胞内の位置: Cytoplasm: P00517
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 43001.87

構造登録者

Bonn, S.,Herrero, S.,Breitenlechner, C.B.,Engh, R.A.,Gassel, M.,Bossemeyer, D. (登録日: 2006-04-10, 公開日: 2006-05-23, 最終更新日: 2024-10-16)

主引用文献

Bonn, S.,Herrero, S.,Breitenlechner, C.B.,Erlbruch, A.,Lehmann, W.,Engh, R.A.,Gassel, M.,Bossemeyer, D.
Structural analysis of protein kinase A mutants with Rho-kinase inhibitor specificity
J.Biol.Chem., 281:24818-24830, 2006

PubMed Abstract: Controlling aberrant kinase-mediated cellular signaling is a major strategy in cancer therapy; successful protein kinase inhibitors such as Tarceva and Gleevec verify this approach. Specificity of inhibitors for the targeted kinase(s), however, is a crucial factor for therapeutic success. Based on homology modeling, we previously identified four amino acids in the active site of Rho-kinase that likely determine inhibitor specificities observed for Rho-kinase relative to protein kinase A (PKA) (in PKA numbering: T183A, L49I, V123M, and E127D), and a fifth (Q181K) that played a surprising role in PKA-PKB hybrid proteins. We have systematically mutated these residues in PKA to their counterparts in Rho-kinase, individually and in combination. Using four Rho-kinase-specific, one PKA-specific, and one pan-kinase-specific inhibitor, we measured the inhibitor-binding properties of the mutated proteins and identify the roles of individual residues as specificity determinants. Two combined mutant proteins, containing the combination of mutations T183A and L49I, closely mimic Rho-kinase. Kinetic results corroborate the hypothesis that side-chain identities form the major determinants of selectivity. An unexpected result of the analysis is the consistent contribution of the individual mutations by simple factors. Crystal structures of the surrogate kinase inhibitor complexes provide a detailed basis for an understanding of these selectivity determinant residues. The ability to obtain kinetic and structural data from these PKA mutants, combined with their Rho-kinase-like selectivity profiles, make them valuable for use as surrogate kinases for structure-based inhibitor design.

PubMed: 16699172
DOI: 10.1074/jbc.M512374200

実験手法

X-RAY DIFFRACTION (1.87 Å)