2LR2

Designed IgG and lanthanide binding probe for solution NMR, MRI and luminescence microscopy

2LR2 の概要

• エントリーDOI: 10.2210/pdb2lr2/pdb
• NMR情報: BMRB: 18126
• 分子名称: Immunoglobulin G-binding protein A (1 entity in total)
• 機能のキーワード: z domain, lanthanide binding tag, de novo protein
• 由来する生物種: artificial gene
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 9875.54

構造登録者

Barb, A.W.,Ho, T.G.,Prestegard, J.H. (登録日: 2012-03-21, 公開日: 2012-09-12, 最終更新日: 2024-05-15)

主引用文献

Barb, A.W.,Ho, T.G.,Flanagan-Steet, H.,Prestegard, J.H.
Lanthanide binding and IgG affinity construct: Potential applications in solution NMR, MRI, and luminescence microscopy.
Protein Sci., 21:1456-1466, 2012

PubMed Abstract: Paramagnetic lanthanide ions when bound to proteins offer great potential for structural investigations that utilize solution nuclear magnetic resonance spectroscopy, magnetic resonance imaging, or optical microscopy. However, many proteins do not have native metal ion binding sites and engineering a chimeric protein to bind an ion while retaining affinity for a protein of interest represents a significant challenge. Here we report the characterization of an immunoglobulin G-binding protein redesigned to include a lanthanide binding motif in place of a loop between two helices (Z-L2LBT). It was shown to bind Tb³⁺ with 130 nM affinity. Ions such as Dy³⁺, Yb³⁺, and Ce³⁺ produce paramagnetic effects on NMR spectra and the utility of these effects is illustrated by their use in determining a structural model of the metal-complexed Z-L2LBT protein and a preliminary characterization of the dynamic distribution of IgG Fc glycan positions. Furthermore, this designed protein is demonstrated to be a novel IgG-binding reagent for magnetic resonance imaging (Z-L2LBT:Gd³⁺ complex) and luminescence microscopy (Z-L2LBT: Tb³⁺ complex).

PubMed: 22851279
DOI: 10.1002/pro.2133

実験手法

SOLUTION NMR