2XIW

Crystal structure of the Sac7d-derived IgG1-binder C3-C24S

2XIW の概要

• エントリーDOI: 10.2210/pdb2xiw/pdb
• 関連するPDBエントリー: 1AZP 1AZQ 1BF4 1CA5 1CA6 1SAP 1WD0 1WD1 1WTO 1WTP 1WTQ 1WTR 1WTV 1WTW 1WTX 1WVL 1XX8 1XYI
• 分子名称: DNA-BINDING PROTEIN 7D, SULFATE ION, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: dna binding protein
• 由来する生物種: SULFOLOBUS ACIDOCALDARIUS
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 18563.65

構造登録者

Bellinzoni, M.,Colinet, S.,Behar, G.,Alzari, P.M.,Pecorari, F. (登録日: 2010-07-01, 公開日: 2011-07-13, 最終更新日: 2024-10-16)

主引用文献

Behar, G.,Bellinzoni, M.,Maillasson, M.,Paillard-Laurance, L.,Alzari, P.M.,He, X.,Mouratou, B.,Pecorari, F.
Tolerance of the Archaeal Sac7D Scaffold Protein to Alternative Library Designs: Characterization of Anti-Immunoglobulin G Affitins.
Protein Eng.Des.Sel., 26:267-, 2013

PubMed Abstract: Engineered protein scaffolds have received considerable attention as alternatives to antibodies in both basic and applied research, as they can offer superior biophysical properties often associated with a simpler molecular organization. Sac7d has been demonstrated as an effective scaffold for molecular recognition. Here, we used the initial L1 'flat surface' library constructed by randomization of 14 residues, to identify ligands specific for human immunoglobulin G. To challenge the plasticity of the Sac7d protein scaffold, we designed the alternative L2 'flat surface & loops' library whereof only 10 residues are randomized. Representative binders (Affitins) of the two libraries exhibited affinities in the low nanomolar range and were able to recognize different epitopes within human immunoglobulin G. These Affitins were stable up to pH 12 while largely conserving other favorable properties of Sac7d protein, such as high expression yields in Escherichia coli, solubility, thermal stability up to 80.7°C, and acidic stability (pH 0). In agreement with our library designs, mutagenesis study revealed two distinct binding areas, one including loops. Together, our results indicate that the Sac7d scaffold tolerates alternative library designs, which further expands the diversity of Affitins and may provide a general way to create tailored affinity tools for demanding applications.

PubMed: 23315487
DOI: 10.1093/PROTEIN/GZS106

実験手法

X-RAY DIFFRACTION (1.5 Å)