5LDF

Maltose binding protein genetically fused to dodecameric glutamine synthetase

5LDF の概要

• エントリーDOI: 10.2210/pdb5ldf/pdb
• EMDBエントリー: 4039
• 関連するBIRD辞書のPRD_ID: PRD_900001
• 分子名称: Glutamine synthetase, Maltose-binding periplasmic protein, alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose (3 entities in total)
• 機能のキーワード: fusion protein, chimera, dodecamer, symmetrized construct, ligase
• 由来する生物種: Salmonella typhi; 詳細
• タンパク質・核酸の鎖数: 24
• 化学式量合計: 1112180.58

構造登録者

Coscia, F.,Petosa, C.,Schoehn, G. (登録日: 2016-06-25, 公開日: 2016-08-10, 最終更新日: 2024-05-15)

主引用文献

Coscia, F.,Estrozi, L.F.,Hans, F.,Malet, H.,Noirclerc-Savoye, M.,Schoehn, G.,Petosa, C.
Fusion to a homo-oligomeric scaffold allows cryo-EM analysis of a small protein.
Sci Rep, 6:30909-30909, 2016

PubMed Abstract: Recent technical advances have revolutionized the field of cryo-electron microscopy (cryo-EM). However, most monomeric proteins remain too small (<100 kDa) for cryo-EM analysis. To overcome this limitation, we explored a strategy whereby a monomeric target protein is genetically fused to a homo-oligomeric scaffold protein and the junction optimized to allow the target to adopt the scaffold symmetry, thereby generating a chimeric particle suitable for cryo-EM. To demonstrate the concept, we fused maltose-binding protein (MBP), a 40 kDa monomer, to glutamine synthetase, a dodecamer formed by two hexameric rings. Chimeric constructs with different junction lengths were screened by biophysical analysis and negative-stain EM. The optimal construct yielded a cryo-EM reconstruction that revealed the MBP structure at sub-nanometre resolution. These findings illustrate the feasibility of using homo-oligomeric scaffolds to enable cryo-EM analysis of monomeric proteins, paving the way for applying this strategy to challenging structures resistant to crystallographic and NMR analysis.

PubMed: 27485862
DOI: 10.1038/srep30909

実験手法

ELECTRON MICROSCOPY (6.2 Å)