8RMO
Crystal structure of anti-FLAG M2 Fab fragment bound to FLAG-tag peptide epitope
Summary for 8RMO
| Entry DOI | 10.2210/pdb8rmo/pdb |
| Related | 7BG1 |
| Descriptor | FLAG-tag, anti-FLAG M2 heavy chain, anti-FLAG M2 light chain, ... (5 entities in total) |
| Functional Keywords | antibody fragment, purification, fab, immune system |
| Biological source | Mus musculus More |
| Total number of polymer chains | 3 |
| Total formula weight | 50022.61 |
| Authors | Beugelink, J.W.,Janssen, B.J.C.,Pronker, M.F. (deposition date: 2024-01-08, release date: 2024-04-03, Last modification date: 2024-10-23) |
| Primary citation | 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 Cited by 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: 38852931DOI: 10.1016/j.jmb.2024.168649 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.163 Å) |
Structure validation
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