4C7N

Crystal Structure of the synthetic peptide iM10 in complex with the coiled-coil region of MITF

4C7N の概要

• エントリーDOI: 10.2210/pdb4c7n/pdb
• 分子名称: MICROPHTHALMIA ASSOCIATED TRANSCRIPTION FACTOR, SYNTHETIC ALPHA-HELIX, IM10, MERCURY (II) ION, ... (4 entities in total)
• 機能のキーワード: transcription, coiled-coil, protein engineering
• 由来する生物種: HOMO SAPIENS (HUMAN); 詳細
• 細胞内の位置: Nucleus: O75030
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 13394.35

構造登録者

Wohlwend, D.,Gerhardt, S.,Kuekenshoener, T.,Einsle, O. (登録日: 2013-09-23, 公開日: 2014-04-02, 最終更新日: 2024-05-08)

主引用文献

Kukenshoner, T.,Wohlwend, D.,Niemoller, J.,Dondapati, P.,Speck, J.,Adeniran, A.V.,Nieth, A.,Gerhardt, S.,Einsle, O.,Muller, K.M.,Arndt, K.M.
Improving Coiled Coil Stability While Maintaining Specificity by a Bacterial Hitchhiker Selection System.
J.Struct.Biol., 186:335-, 2014

PubMed Abstract: The design and selection of peptides targeting cellular proteins is challenging and often yields candidates with undesired properties. Therefore we deployed a new selection system based on the twin-arginine translocase (TAT) pathway of Escherichia coli, named hitchhiker translocation (HiT) selection. A pool of α-helix encoding sequences was designed and selected for interference with the coiled coil domain (CC) of a melanoma-associated basic-helix-loop-helix-leucine-zipper (bHLHLZ) protein, the microphthalmia associated transcription factor (MITF). One predominant sequence (iM10) was enriched during selection and showed remarkable protease resistance, high solubility and thermal stability while maintaining its specificity. Furthermore, it exhibited nanomolar range affinity towards the target peptide. A mutation screen indicated that target-binding helices of increased homodimer stability and improved expression rates were preferred in the selection process. The crystal structure of the iM10/MITF-CC heterodimer (2.1Å) provided important structural insights and validated our design predictions. Importantly, iM10 did not only bind to the MITF coiled coil, but also to the markedly more stable HLHLZ domain of MITF. Characterizing the selected variants of the semi-rational library demonstrated the potential of the innovative bacterial selection approach.

PubMed: 24631970
DOI: 10.1016/J.JSB.2014.03.002

実験手法

X-RAY DIFFRACTION (2.1 Å)