Engineered nanobodies facilitate cryo-EM studies of small proteins

Subjects

Single-particle cryo-electron microscopy (cryo-EM) has become a conventional approach in structural biology, but resolving the structures of small proteins (<100 kDa) presents substantial challenges, mainly owing to low signal-to-noise ratios. The resulting difficulty in particle picking and alignment during data processing produces inaccuracies, ultimately leading to greater uncertainty in density locations and lower resolutions for 3D reconstructions. To overcome this limitation, we have developed an engineered-nanobody strategy, offering a promising and potentially universal solution to this bottleneck.

The Di-Gembody scaffold serves as a model of modular design and functional adaptability, coupled with an efficient workflow for structure determination without extensive protein engineering. This approach considerably augments the cryo-EM methodological repertoire by incorporating a strategically designed nanobody interface and creating quaternary structures through chemical alteration, ultimately providing valuable insights that may inform the development of nanobody techniques across a wide range of biotechnological applications.

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