| Autor: |
Lu M; Department of Chemistry and Biochemistry , University of Delaware , 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.; Department of Structural Biology , University of Pittsburgh School of Medicine , 3501 Fifth Avenue , Pittsburgh , Pennsylvania 15261 , United States., Wang M; Department of Chemistry and Biochemistry , University of Delaware , 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., Sergeyev IV; Bruker Biospin Corporation , 15 Fortune Drive , Billerica , Massachusetts 01821 , United States., Quinn CM; Department of Chemistry and Biochemistry , University of Delaware , Newark , Delaware 19716 , United States., Struppe J; Bruker Biospin Corporation , 15 Fortune Drive , Billerica , Massachusetts 01821 , United States., Rosay M; Bruker Biospin Corporation , 15 Fortune Drive , Billerica , Massachusetts 01821 , United States., Maas W; Bruker Biospin Corporation , 15 Fortune Drive , Billerica , Massachusetts 01821 , 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; Department of Chemistry and Biochemistry , University of Delaware , 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. |
| Abstrakt: |
We report remarkably high, up to 100-fold, signal enhancements in 19 F dynamic nuclear polarization (DNP) magic angle spinning (MAS) spectra at 14.1 T on HIV-1 capsid protein (CA) assemblies. These enhancements correspond to absolute sensitivity ratios of 12-29 and are of similar magnitude to those seen for 1 H signals in the same samples. At MAS frequencies above 20 kHz, it was possible to record 2D 19 F- 13 C HETCOR spectra, which contain long-range intra- and intermolecular correlations. Such correlations provide unique distance restraints, inaccessible in conventional experiments without DNP, for protein structure determination. Furthermore, systematic quantification of the DNP enhancements as a function of biradical concentration, MAS frequency, temperature, and microwave power is reported. Our work establishes the power of DNP-enhanced 19 F MAS NMR spectroscopy for structural characterization of HIV-1 CA assemblies, and this approach is anticipated to be applicable to a wide range of large biomolecular systems. |
