Integrating 19 F Distance Restraints for Accurate Protein Structure Determination by Magic Angle Spinning NMR Spectroscopy.

Autor: Runge BR; University of Delaware, Department of Chemistry and Biochemistry, Newark, Delaware 19716, United States.; Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, 1051 Biomedical Science Tower 3, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States., Zadorozhnyi R; University of Delaware, Department of Chemistry and Biochemistry, Newark, Delaware 19716, United States.; Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, 1051 Biomedical Science Tower 3, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States., Quinn CM; University of Delaware, Department of Chemistry and Biochemistry, Newark, Delaware 19716, United States., Russell RW; University of Delaware, Department of Chemistry and Biochemistry, Newark, Delaware 19716, United States.; Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, 1051 Biomedical Science Tower 3, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States., Lu M; Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, 1051 Biomedical Science Tower 3, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States.; Department of Structural Biology, University of Pittsburgh School of Medicine, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States., Antolínez S; University of Delaware, Department of Chemistry and Biochemistry, Newark, Delaware 19716, United States., Struppe J; Bruker Biospin Corporation, 15 Fortune Drive, Billerica, Massachusetts 01821, United States., Schwieters CD; Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 12A, Bethesda, Maryland 20892, United States., Byeon IL; Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, 1051 Biomedical Science Tower 3, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States.; Department of Structural Biology, University of Pittsburgh School of Medicine, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States., Hadden-Perilla JA; University of Delaware, Department of Chemistry and Biochemistry, Newark, Delaware 19716, United States., Gronenborn AM; Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, 1051 Biomedical Science Tower 3, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States.; Department of Structural Biology, University of Pittsburgh School of Medicine, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States., Polenova T; University of Delaware, Department of Chemistry and Biochemistry, Newark, Delaware 19716, United States.; Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, 1051 Biomedical Science Tower 3, 3501 Fifth Avenue, Pittsburgh, Pennsylvania 15261, United States.
Jazyk: angličtina
Zdroj: Journal of the American Chemical Society [J Am Chem Soc] 2024 Nov 06; Vol. 146 (44), pp. 30483-30494. Date of Electronic Publication: 2024 Oct 23.
DOI: 10.1021/jacs.4c11373
Abstrakt: Traditional protein structure determination by magic angle spinning (MAS) solid-state NMR spectroscopy primarily relies on interatomic distances up to 8 Å, extracted from 13 C-, 15 N-, and 1 H-based dipolar-based correlation experiments. Here, we show that 19 F fast (60 kHz) MAS NMR spectroscopy can supply additional, longer distances. Using 4F-Trp,U- 13 C, 15 N crystalline Oscillatoria agardhii agglutinin (OAA), we demonstrate that judiciously designed 2D and 3D 19 F-based dipolar correlation experiments such as (H)CF, (H)CHF, and FF can yield interatomic distances in the 8-16 Å range. Incorporation of fluorine-based restraints into structure calculation improved the precision of Trp side chain conformations as well as regions in the protein around the fluorine containing residues, with notable improvements observed for residues in proximity to the Trp pairs (W10/W17 and W77/W84) in the carbohydrate-binding loops, which lacked sufficient long-range 13 C- 13 C distance restraints. Our work highlights the use of fluorine and 19 F fast MAS NMR spectroscopy as a powerful structural biology tool.
Databáze: MEDLINE
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