9JQE
Cryo-EM structure of ferritin variant R63MeH/R67MeH with Cu(II)
Summary for 9JQE
| Entry DOI | 10.2210/pdb9jqe/pdb |
| EMDB information | 61729 |
| Descriptor | Ferritin heavy chain, COPPER (II) ION (3 entities in total) |
| Functional Keywords | cryo-em, ferritin variant, non-canonical amino acid, h-n-3-methyl-l-histidine(meh), metal binding protein |
| Biological source | Homo sapiens (human) |
| Total number of polymer chains | 24 |
| Total formula weight | 513570.53 |
| Authors | Wang, C.H.,Wang, Y.S. (deposition date: 2024-09-27, release date: 2024-11-27, Last modification date: 2024-12-25) |
| Primary citation | Tsou, J.C.,Tsou, C.J.,Wang, C.H.,Ko, A.A.,Wang, Y.H.,Liang, H.H.,Sun, J.C.,Huang, K.F.,Ko, T.P.,Lin, S.Y.,Wang, Y.S. Site-Specific Histidine Aza-Michael Addition in Proteins Enabled by a Ferritin-Based Metalloenzyme. J.Am.Chem.Soc., 146:33309-33315, 2024 Cited by PubMed Abstract: Histidine modifications of proteins are broadly based on chemical methods triggering N-substitution reactions such as aza-Michael addition at histidine's moderately nucleophilic imidazole side chain. While recent studies have demonstrated chemoselective, histidine-specific modifications by further exploiting imidazole's electrophilic reactivity to overcome interference from the more nucleophilic lysine and cysteine, achieving site-specific histidine modifications remains a major challenge due to the absence of spatial control over chemical processes. Herein, through X-ray crystallography and cryo-electron microscopy structural studies, we describe the rational design of a nature-inspired, noncanonical amino-acid-incorporated, human ferritin-based metalloenzyme that is capable of introducing site-specific post-translational modifications (PTMs) to histidine in peptides and proteins. Specifically, chemoenzymatic aza-Michael additions on single histidine residues were carried out on eight protein substrates ranging from 10 to 607 amino acids including the insulin peptide hormone. By introducing an insulin-targeting peptide into our metalloenzyme, we further directed modifications to be carried out site-specifically on insulin's B-chain histidine 5. The success of this biocatalysis platform outlines a novel approach in introducing residue- and, moreover, site-specific post-translational modifications to peptides and proteins, which may further enable reactions to be carried out . PubMed: 39499210DOI: 10.1021/jacs.4c14446 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (1.83 Å) |
Structure validation
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