6XUX

Crystal structure of Megabody Mb-Nb207-cYgjK_NO

6XUX の概要

• エントリーDOI: 10.2210/pdb6xux/pdb
• 分子名称: Nanobody,Glucosidase YgjK,Glucosidase YgjK,Nanobody, CALCIUM ION (3 entities in total)
• 機能のキーワード: megabody, scaffold, structural protein
• 由来する生物種: Escherichia coli K-12; 詳細
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 101659.08

構造登録者

Steyaert, J.,Uchanski, T.,Fischer, B. (登録日: 2020-01-21, 公開日: 2021-01-13, 最終更新日: 2024-10-23)

主引用文献

Uchanski, T.,Masiulis, S.,Fischer, B.,Kalichuk, V.,Lopez-Sanchez, U.,Zarkadas, E.,Weckener, M.,Sente, A.,Ward, P.,Wohlkonig, A.,Zogg, T.,Remaut, H.,Naismith, J.H.,Nury, H.,Vranken, W.,Aricescu, A.R.,Pardon, E.,Steyaert, J.
Megabodies expand the nanobody toolkit for protein structure determination by single-particle cryo-EM.
Nat.Methods, 18:60-68, 2021

PubMed Abstract: Nanobodies are popular and versatile tools for structural biology. They have a compact single immunoglobulin domain organization, bind target proteins with high affinities while reducing their conformational heterogeneity and stabilize multi-protein complexes. Here we demonstrate that engineered nanobodies can also help overcome two major obstacles that limit the resolution of single-particle cryo-electron microscopy reconstructions: particle size and preferential orientation at the water-air interfaces. We have developed and characterized constructs, termed megabodies, by grafting nanobodies onto selected protein scaffolds to increase their molecular weight while retaining the full antigen-binding specificity and affinity. We show that the megabody design principles are applicable to different scaffold proteins and recognition domains of compatible geometries and are amenable for efficient selection from yeast display libraries. Moreover, we demonstrate that megabodies can be used to obtain three-dimensional reconstructions for membrane proteins that suffer from severe preferential orientation or are otherwise too small to allow accurate particle alignment.

PubMed: 33408403
DOI: 10.1038/s41592-020-01001-6

実験手法

X-RAY DIFFRACTION (1.90000643078 Å)