4C59

Structure of GAK kinase in complex with nanobody (NbGAK_4)

4C59 の概要

• エントリーDOI: 10.2210/pdb4c59/pdb
• 関連するPDBエントリー: 4C57 4C58
• 分子名称: Cyclin-G-associated kinase, NANOBODY, (2Z,3E)-2,3'-BIINDOLE-2',3(1H,1'H)-DIONE 3-{O-[(3R)-3,4-DIHYDROXYBUTYL]OXIME}, ... (4 entities in total)
• 機能のキーワード: transferase, kinase, conformational plasticity, activation
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 53634.44

構造登録者

Chaikuad, A.,Keates, T.,Allerston, C.K.,Gileadi, O.,von Delft, F.,Arrowsmith, C.H.,Edwards, A.M.,Bountra, C.,Knapp, S.,Muller-Knapp, S. (登録日: 2013-09-10, 公開日: 2013-10-09, 最終更新日: 2024-10-16)

主引用文献

Chaikuad, A.,Keates, T.,Vincke, C.,Kaufholz, M.,Zenn, M.,Zimmermann, B.,Gutierrez, C.,Zhang, R.G.,Hatzos-Skintges, C.,Joachimiak, A.,Muyldermans, S.,Herberg, F.W.,Knapp, S.,Muller, S.
Structure of cyclin G-associated kinase (GAK) trapped in different conformations using nanobodies.
Biochem. J., 459:59-69, 2014

PubMed Abstract: GAK (cyclin G-associated kinase) is a key regulator of clathrin-coated vesicle trafficking and plays a central role during development. Additionally, due to the unusually high plasticity of its catalytic domain, it is a frequent 'off-target' of clinical kinase inhibitors associated with respiratory side effects of these drugs. In the present paper, we determined the crystal structure of the GAK catalytic domain alone and in complex with specific single-chain antibodies (nanobodies). GAK is constitutively active and weakly associates in solution. The GAK apo structure revealed a dimeric inactive state of the catalytic domain mediated by an unusual activation segment interaction. Co-crystallization with the nanobody NbGAK_4 trapped GAK in a dimeric arrangement similar to the one observed in the apo structure, whereas NbGAK_1 captured the activation segment of monomeric GAK in a well-ordered conformation, representing features of the active kinase. The presented structural and biochemical data provide insight into the domain plasticity of GAK and demonstrate the utility of nanobodies to gain insight into conformational changes of dynamic molecules. In addition, we present structural data on the binding mode of ATP mimetic inhibitors and enzyme kinetic data, which will support rational inhibitor design of inhibitors to reduce the off-target effect on GAK.

PubMed: 24438162
DOI: 10.1042/BJ20131399

実験手法

X-RAY DIFFRACTION (2.8 Å)