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Публикация

Kaitlyn Abe

1/5: Our latest preprint is out! We show that LEA proteins can diversify particle orientations in cryo-EM, helping to overcome preferred orientation at the air-water interface (AWI). www.biorxiv.org/cgi/content/... with @cijilim.bsky.social and Tim Grant #CryoEM #LEAproteins

Diversifying particle-orientation distributions in cryo-EM with LEA protein additivesPreferential orientation at the air-water interface (AWI) remains a persistent challenge in cryo-electron microscopy, often requiring extensive optimization or specialized grid preparation strategies....www.biorxiv.org
Обсуждение

1 прямых ответов · 4 сообщений

Kaitlyn Abe

2/5: We've previously shown that LEA proteins AavLEA1 and RvLEAMshort can protect fragile samples from AWI damage, but we've found that these proteins can also alter sample orientation distributions.

Ответ для Kaitlyn Abe
Kaitlyn Abe

3/5: HA trimer is a benchmark for severe preferred orientation. Adding LEA proteins dramatically broadened angular distribution coverage and improved map quality without relying on a stage-tilted data collection.

Ответ для Kaitlyn Abe
Kaitlyn Abe

4/5: We've also explanded from two to six LEA proteins. All six protected Polα-primase from AWI damage but each produced a unique orientation distribution suggesting that AWI protection is a shared property while orientation control is LEA-specific.

Ответ для Kaitlyn Abe
Kaitlyn Abe

5/5: Combining datasets collected with different LEA proteins computationally produced more isotropic reconstructions. Surprisingly, premixing multiple LEA proteins before grid preparation did not simply combine their effects and produced unexpected orientation distributions.