Megabodies expand the nanobody toolkit for protein structure determination by single-particle cryo-EM.

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 diff

SUBMITTER: Uchanski T 

PROVIDER: S-EPMC7611088 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature