6C4U

Engineered FHA with Myc-pTBD peptide

6C4U の概要

• エントリーDOI: 10.2210/pdb6c4u/pdb
• 分子名称: Forkhead-associated 1, Myc-pTBD peptide, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: fha, protein engineering, myc pt58 target, peptide binding protein
• 由来する生物種: Saccharomyces cerevisiae (baker's yeast); 詳細
• タンパク質・核酸の鎖数: 12
• 化学式量合計: 97173.98

構造登録者

Kall, S.L.,Lavie, A. (登録日: 2018-01-12, 公開日: 2018-05-30, 最終更新日: 2024-11-13)

主引用文献

Venegas, L.A.,Kall, S.L.,Bankole, O.,Lavie, A.,Kay, B.K.
Generating a recombinant phosphothreonine-binding domain for a phosphopeptide of the human transcription factor, c-Myc.
N Biotechnol, 45:36-44, 2018

PubMed Abstract: Transcription factor c-Myc is an oncoprotein that is regulated at the post-translational level through phosphorylation of two conserved residues, Serine 62 (Ser62) and Threonine 58 (Thr58). A highly specific tool capable of recognizing Myc via pThr58 is needed to monitor activation and localization. Through phage display, we have isolated 10 engineered Forkhead-associated (FHA) domains that selectively bind to a phosphothreonine (pThr)-containing peptide (53-FELLPpTPPLSPS-64) segment of human c-Myc. One domain variant was observed to bind to the Myc-pThr58 peptide with a K value of 800 nM and had >1000-fold discrimination between the phosphorylated and non-phosphorylated peptide. The crystal structure of the engineered FHA Myc-pThr-binding domain (Myc-pTBD) was solved in complex with its cognate ligand. The Myc-pTBD was observed to be structurally similar to the yeast Rad9 FHA1 domain, except that its β4-β5 and β10-β11 loops form a hydrophobic pocket to facilitate the interaction between the domain and the peptide ligand. The Myc-pTBD's specificity for its cognate ligand was demonstrated to be on a par with 3 commercial polyclonal antibodies, suggesting that this recombinant reagent is a viable alternative to antibodies for monitoring Myc regulation.

PubMed: 29763736
DOI: 10.1016/j.nbt.2018.05.001

実験手法

X-RAY DIFFRACTION (2.6 Å)