5OBB

Structure of a modified mouse H chain ferritin with a lanthanide binding motif in complex with Terbium

5OBB の概要

• エントリーDOI: 10.2210/pdb5obb/pdb
• 分子名称: Ferritin heavy chain, TERBIUM(III) ION (3 entities in total)
• 機能のキーワード: ferritin, lanthanide, terbium, metal transport
• 由来する生物種: Mus musculus (Mouse)
• タンパク質・核酸の鎖数: 24
• 化学式量合計: 549701.62

構造登録者

Baiocco, P.,Trabuco, M.C. (登録日: 2017-06-26, 公開日: 2018-07-25, 最終更新日: 2024-01-17)

主引用文献

Calisti, L.,Trabuco, M.C.,Boffi, A.,Testi, C.,Montemiglio, L.C.,des Georges, A.,Benni, I.,Ilari, A.,Taciak, B.,Bialasek, M.,Rygiel, T.,Krol, M.,Baiocco, P.,Bonamore, A.
Engineered ferritin for lanthanide binding.
PLoS ONE, 13:e0201859-e0201859, 2018

PubMed Abstract: Ferritin H-homopolymers have been extensively used as nanocarriers for diverse applications in the targeted delivery of drugs and imaging agents, due to their unique ability to bind the transferrin receptor (CD71), highly overexpressed in most tumor cells. In order to incorporate novel fluorescence imaging properties, we have fused a lanthanide binding tag (LBT) to the C-terminal end of mouse H-chain ferritin, HFt. The HFt-LBT possesses one high affinity Terbium binding site per each of the 24 subunits provided by six coordinating aminoacid side chains and a tryptophan residue in its close proximity and is thus endowed with strong FRET sensitization properties. Accordingly, the characteristic Terbium emission band at 544 nm for the HFt-LBT Tb(III) complex was detectable upon excitation of the tag enclosed at two order of magnitude higher intensity with respect to the wtHFt protein. X-ray data at 2.9 Å and cryo-EM at 7 Å resolution demonstrated that HFt-LBT is correctly assembled as a 24-mer both in crystal and in solution. On the basis of the intrinsic Tb(III) binding properties of the wt protein, 32 additional Tb(III) binding sites, located within the natural iron binding sites of the protein, were identified besides the 24 Tb(III) ions coordinated to the LBTs. HFt-LBT Tb(III) was demonstrated to be actively uptaken by selected tumor cell lines by confocal microscopy and FACS analysis of their FITC derivatives, although direct fluorescence from Terbium emission could not be singled out with conventional, 295-375 nm, fluorescence excitation.

PubMed: 30102720
DOI: 10.1371/journal.pone.0201859

実験手法

X-RAY DIFFRACTION (2.65 Å)