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| Title | Quasi-continuous cotranslational compaction and folding of a multidomain protein. |
|---|---|
| Journal, issue, pages | bioRxiv, Year 2026 |
| Publish date | Feb 6, 2026 |
 Authors | Spyridoula Mitsikosta / Justin M Westerfield / Fátima Pardo-Avila / Michael Levitt / Gunnar von Heijne / Ane Metola /   |
| PubMed Abstract | Most proteins start to fold cotranslationally as they come off the ribosome. So far, studies of cotranslational folding have focused mainly on small, single-domain proteins. Here, we have used Force ...Most proteins start to fold cotranslationally as they come off the ribosome. So far, studies of cotranslational folding have focused mainly on small, single-domain proteins. Here, we have used Force Profile Analysis to study the cotranslational folding of Firefly luciferase, a complex 550-residue protein composed of an N-terminal domain (NTD) encompassing two split Rossmann folds (RF-1, RF-2) and a β-roll, and a flexibly attached C-terminal domain (CTD). The folding process is characterized by a quasi-continuous series of compaction/folding steps that generate intermediate-size pulling forces on the nascent chain, punctuated by a prominent high-force event that represents the folding of the RF-2 domain, and a few low-force instances that likely indicate the formation of distinct folding intermediates. Trigger Factor interacts extensively with the nascent chain when the central part of RF-2 and the early parts of the CTD are synthesized. Our analysis uncovers a cotranslational compaction/folding process that is rich in detail and not just a simple succession of a few distinct, cooperative folding transitions. |
 External links | bioRxiv / PubMed:41676707 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 2.14 - 2.29 Å |
| Structure data |  EMDB-56368: Escherichia coli ribosome arrested on a chimeric, 110-residue construct featuring a firefly-luciferase truncation followed by SecM(3W)  EMDB-56453: Escherichia coli ribosome arrested on a chimeric, 130-residue construct featuring a firefly-luciferase truncation followed by SecM(3W)  EMDB-56462: Escherichia coli ribosome arrested on a chimeric, 190-residue construct featuring a firefly-luciferase truncation followed by SecM(3W) |
| Source |
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Escherichia coli BL21(DE3) (bacteria)