Journal
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS
Volume 75, Issue -, Pages 547-551Publisher
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S2053230X19010136
Keywords
RNA structure; fluorescence turn-on aptamers; X-ray free-electron lasers; iMango-III; fluorescence; XFELS; room temperature
Funding
- National Cancer Institute
- National Heart, Lung and Blood Institute, NIH
- NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [ZIAHL006188] Funding Source: NIH RePORTER
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Turn-on aptamers are in vitro-selected RNAs that bind to conditionally fluorescent small molecules and enhance their fluorescence. Upon binding TO1-biotin, the iMango-III aptamer achieves the largest fluorescence enhancement reported for turn-on aptamers (over 5000-fold). This aptamer was generated by structure-guided engineering and functional reselection of the parental aptamer Mango-III. Structures of both Mango-III and iMango-III have previously been determined by conventional cryocrystallography using synchrotron X-radiation. Using an X-ray free-electron laser (XFEL), the room-temperature iMango-III-TO1-biotin co-crystal structure has now been determined at 3.0 angstrom resolution. This structural model, which was refined against a data set of similar to 1300 diffraction images (each from a single crystal), is largely consistent with the structures determined from single-crystal data sets collected at 100 K. This constitutes a technical benchmark on the way to XFEL pump-probe experiments on fluorescent RNA-small molecule complexes.
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