5XHM

Crystal structure of Frog M-ferritin D40A mutant

5XHM の概要

• エントリーDOI: 10.2210/pdb5xhm/pdb
• 関連するPDBエントリー: 5XHI 5XHN 5XHO
• 分子名称: Ferritin, middle subunit, MAGNESIUM ION, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: ferritin, m-ferritin, oxidoreductase
• 由来する生物種: Rana catesbeiana (American bullfrog)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 20887.57

構造登録者

Jagdev, M.K.,Vasudevan, D. (登録日: 2017-04-21, 公開日: 2017-08-09, 最終更新日: 2023-11-22)

主引用文献

Subhadarshanee, B.,Mohanty, A.,Jagdev, M.K.,Vasudevan, D.,Behera, R.K.
Surface charge dependent separation of modified and hybrid ferritin in native PAGE: Impact of lysine 104
Biochim. Biophys. Acta, 1865:1267-1273, 2017

PubMed Abstract: Preparation of modified and hybrid ferritin provides a great opportunity to understand the mechanisms of iron loading/unloading, protein self-assembly, size constrained nanomaterial synthesis and targeted drug delivery. However, the large size (M.W.=490kDa) has been limiting the separation of different modified and/or hybrid ferritin nanocages from each other in their intact assembled form and further characterization. Native polyacrylamide gel electrophoresis (PAGE) separates proteins on the basis of both charge and mass, while maintaining their overall native structure and activity. Altering surface charge distribution by substitution of amino acid residues located at the external surface of ferritin (K104E & D40A) affected the migration rate in native PAGE while internal modification had little effect. Crystal structures confirmed that ferritin nanocages made up of subunits with single amino acid substitutions retain the overall structure of ferritin nanocage. Taking advantage of K104E migration behavior, formation of hybrid ferritins with subunits of wild type (WT) and K104E were confirmed and separated in native PAGE. Cage integrity and iron loading ability (ferritin activity) were also tested. The migration pattern of hybrid ferritins in native PAGE depends on the subunit ratio (WT: K104E) in the ferritin cage. Our work shows that native PAGE can be exploited in nanobiotechnology, by analyzing modifications of large proteins like ferritin.

PubMed: 28739445
DOI: 10.1016/j.bbapap.2017.07.012

実験手法

X-RAY DIFFRACTION (1.7 Å)