8RMO

Crystal structure of anti-FLAG M2 Fab fragment bound to FLAG-tag peptide epitope

8RMO の概要

• エントリーDOI: 10.2210/pdb8rmo/pdb
• 関連するPDBエントリー: 7BG1
• 分子名称: FLAG-tag, anti-FLAG M2 heavy chain, anti-FLAG M2 light chain, ... (5 entities in total)
• 機能のキーワード: antibody fragment, purification, fab, immune system
• 由来する生物種: Mus musculus; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 50022.61

構造登録者

Beugelink, J.W.,Janssen, B.J.C.,Pronker, M.F. (登録日: 2024-01-08, 公開日: 2024-04-03, 最終更新日: 2024-10-23)

主引用文献

Beugelink, J.W.,Sweep, E.,Janssen, B.J.C.,Snijder, J.,Pronker, M.F.
Structural Basis for Recognition of the FLAG-tag by Anti-FLAG M2.
J.Mol.Biol., 436:168649-168649, 2024

PubMed Abstract: The FLAG-tag/anti-FLAG system is a widely used biochemical tool for protein detection and purification. Anti-FLAG M2 is the most popular antibody against the FLAG-tag, due to its ease of use, versatility, and availability in pure form or as bead conjugate. M2 binds N-terminal, C-terminal and internal FLAG-tags and binding is calcium-independent, but the molecular basis for the FLAG-tag specificity and recognition remains unresolved. Here we present an atomic resolution (1.17 Å) structure of the FLAG peptide in complex with the Fab of anti-FLAG M2, revealing key binding determinants. Five of the eight FLAG peptide residues form direct interactions with paratope residues. The FLAG peptide adopts a 3 helix conformation in complex with the Fab. These structural insights allowed us to rationally introduce point mutations on both the peptide and antibody side. We tested these by surface plasmon resonance, leading us to propose a shorter yet equally binding version of the FLAG-tag for the M2 antibody.

PubMed: 38852931
DOI: 10.1016/j.jmb.2024.168649

実験手法

X-RAY DIFFRACTION (1.163 Å)