9JQB

Cryo-EM structure of ferritin variant R63BrThA/E67BrThA

9JQB の概要

• エントリーDOI: 10.2210/pdb9jqb/pdb
• EMDBエントリー: 61726
• 分子名称: Ferritin heavy chain, SODIUM ION (3 entities in total)
• 機能のキーワード: cryo-em, ferritin variant, non-canonical amino acid, l-2-(5-bromothienyl)alanine (brtha), metal binding protein
• 由来する生物種: Homo sapiens (human)
• タンパク質・核酸の鎖数: 24
• 化学式量合計: 514956.84

構造登録者

Wang, C.H.,Sun, J.C.,Wang, Y.S. (登録日: 2024-09-27, 公開日: 2024-11-27, 最終更新日: 2024-12-25)

主引用文献

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

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: 39499210
DOI: 10.1021/jacs.4c14446

実験手法

ELECTRON MICROSCOPY (1.78 Å)