2BKK

Crystal structure of Aminoglycoside Phosphotransferase APH(3')-IIIa in complex with the inhibitor AR_3a

2BKK の概要

• エントリーDOI: 10.2210/pdb2bkk/pdb
• 関連するPDBエントリー: 1J7I 1J7L 1J7U 1K1A 1K1B 1L8T 1L8U 1MJ0 1N0Q 1N0R 2BKG
• 分子名称: AMINOGLYCOSIDE 3'-PHOSPHOTRANSFERASE, DESIGNED ANKYRIN REPEAT INHIBITOR AR_3A, ADENOSINE-5'-DIPHOSPHATE, ... (5 entities in total)
• 機能のキーワード: transferase/peptide, transferase-designed protein complex, ankyrin repeat, co-crystallization, inhibitor design, drug design, enzyme inhibition, kinase inhibition, designed repeat protein, antibiotic resistance, atp-binding, kinase, plasmid, transferase, transferase-peptide complex
• 由来する生物種: ENTEROCOCCUS FAECALIS; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 100117.13

構造登録者

Kohl, A.,Amstutz, P.,Parizek, P.,Binz, H.K.,Briand, C.,Capitani, G.,Forrer, P.,Pluckthun, A.,Grutter, M.G. (登録日: 2005-02-16, 公開日: 2005-08-09, 最終更新日: 2023-12-13)

主引用文献

Kohl, A.,Amstutz, P.,Parizek, P.,Binz, H.K.,Briand, C.,Capitani, G.,Forrer, P.,Pluckthun, A.,Grutter, M.G.
Allosteric Inhibition of Aminoglycoside Phosphotransferase by a Designed Ankyrin Repeat Protein
Structure, 13:1131-, 2005

PubMed Abstract: Aminoglycoside phosphotransferase (3')-IIIa (APH) is a bacterial kinase that confers antibiotic resistance to many pathogenic bacteria and shares structural homology with eukaryotic protein kinases. We report here the crystal structure of APH, trapped in an inactive conformation by a tailor-made inhibitory ankyrin repeat (AR) protein, at 2.15 A resolution. The inhibitor was selected from a combinatorial library of designed AR proteins. The AR protein binds the C-terminal lobe of APH and thereby stabilizes three alpha helices, which are necessary for substrate binding, in a significantly displaced conformation. BIAcore analysis and kinetic enzyme inhibition experiments are consistent with the proposed allosteric inhibition mechanism. In contrast to most small-molecule kinase inhibitors, the AR proteins are not restricted to active site binding, allowing for higher specificity. Inactive conformations of pharmaceutically relevant enzymes, as can be elucidated with the approach presented here, represent powerful starting points for rational drug design.

PubMed: 16084385
DOI: 10.1016/J.STR.2005.04.020

実験手法

X-RAY DIFFRACTION (2.15 Å)