9VX2
Crystal structure of the apo form of HisMab-1 Fab
Summary for 9VX2
| Entry DOI | 10.2210/pdb9vx2/pdb |
| Descriptor | HisMab-1 immunoglobulin heavy chain variable region, HisMab-1 immunoglobulin light chain variable region, SULFATE ION, ... (4 entities in total) |
| Functional Keywords | fab, hismab-1, his-tag, immune system |
| Biological source | Mus musculus More |
| Total number of polymer chains | 4 |
| Total formula weight | 96979.25 |
| Authors | Hoshi, S.,Arimori, T.,Harada-Hikita, A. (deposition date: 2025-07-18, release date: 2025-12-17, Last modification date: 2025-12-24) |
| Primary citation | Hitomi, N.,Hoshi, S.,Kaneko, M.K.,Kato, R.,Iwasaki, K.,Takagi, J.,Kato, Y.,Harada-Hikita, A.,Arimori, T. Functional and Structural Characterization of a Novel Anti-His-tag Antibody, HisMab-1. J.Mol.Biol., 438:169574-169574, 2025 Cited by PubMed Abstract: The polyhistidine tag (His-tag) is one of the most widely used peptide tags for the purification of recombinant proteins, owing to its compatibility with immobilized metal affinity chromatography. While numerous anti-His-tag antibodies are commercially available, their quantitative affinity data and structural insights are limited. Here, we present a detailed physicochemical and structural characterization of a novel anti-His-tag antibody, HisMab-1. Isothermal titration calorimetry showed that the Fab fragment of HisMab-1 binds to a hexahistidine peptide in an enthalpy-driven manner, with a dissociation constant (K) of ∼30 nM at a neutral pH. The crystal structure of the HisMab-1-hexahistidine peptide complex at 2.39-Å resolution revealed that HisMab-1 primarily recognizes the first, second, fourth, and fifth histidine residues of the peptide through multiple interactions, including hydrogen bonding and π-π stacking, which collectively contribute to the high specificity of the antibody. Notably, HisMab-1 also binds to a His-tag embedded within a conformationally constrained β-hairpin loop without reducing affinity, highlighting its structural adaptability. These findings establish HisMab-1 as a high-affinity, high-specificity, structurally validated anti-His-tag antibody with broad potential in diverse protein engineering and structural biology applications. PubMed: 41349762DOI: 10.1016/j.jmb.2025.169574 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.39 Å) |
Structure validation
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