Limits...
Novel Chemical Ligands to Ebola Virus and Marburg Virus Nucleoproteins Identified by Combining Affinity Mass Spectrometry and Metabolomics Approaches.

Fu X, Wang Z, Li L, Dong S, Li Z, Jiang Z, Wang Y, Shui W - Sci Rep (2016)

Bottom Line: Accompanying biophysical analyses demonstrate that binding of 18β-glycyrrhetinic acid to EBOV NP significantly reduces protein thermal stability, induces formation of large NP oligomers, and disrupts the critical association of viral ssRNA with NP complexes whereas the compound showed no such activity on MARV NP.Our study has revealed the substantial potential of new analytical techniques in ligand discovery from natural herb resources.In addition, identification of a chemical ligand that influences the oligomeric state and RNA-binding function of EBOV NP sheds new light on antiviral drug development.

View Article: PubMed Central - PubMed

Affiliation: College of Biotechnology, Tianjin University of Science &Technology, Tianjin 300457, China.

ABSTRACT
The nucleoprotein (NP) of Ebola virus (EBOV) and Marburg virus (MARV) is an essential component of the viral ribonucleoprotein complex and significantly impacts replication and transcription of the viral RNA genome. Although NP is regarded as a promising antiviral druggable target, no chemical ligands have been reported to interact with EBOV NP or MARV NP. We identified two compounds from a traditional Chinese medicine Gancao (licorice root) that can bind both NPs by combining affinity mass spectrometry and metabolomics approaches. These two ligands, 18β-glycyrrhetinic acid and licochalcone A, were verified by defined compound mixture screens and further characterized with individual ligand binding assays. Accompanying biophysical analyses demonstrate that binding of 18β-glycyrrhetinic acid to EBOV NP significantly reduces protein thermal stability, induces formation of large NP oligomers, and disrupts the critical association of viral ssRNA with NP complexes whereas the compound showed no such activity on MARV NP. Our study has revealed the substantial potential of new analytical techniques in ligand discovery from natural herb resources. In addition, identification of a chemical ligand that influences the oligomeric state and RNA-binding function of EBOV NP sheds new light on antiviral drug development.

No MeSH data available.


Related in: MedlinePlus

Structural elucidation of GC7 (A) and GC13 (B) by MS and MSMS analysis.The proposed fragmentation pathway is shown below mass spectra.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4940736&req=5

f2: Structural elucidation of GC7 (A) and GC13 (B) by MS and MSMS analysis.The proposed fragmentation pathway is shown below mass spectra.

Mentions: Among the identified licorice constituents, we found GC7/10 (glycyrrhetinic acid) and GC13 (licochalcone A) met the aforementioned thresholds for both EBOV NP and MARV NP ligands (red symbols in Fig. 1C). It is noteworthy that GC7 (18β-glycyrrhetinic acid) and GC10 (18α-glycyrrhetinic acid) are a pair of diastereo-isomers differing only in their C18–H–, trans-, and cis-configuration, and they coeluted under the LC condition of this study. High-resolution MS and MSMS spectra were acquired on GC7/10 and GC13 in the protein incubation sample to support structural elucidation (Fig. 2). In addition, NMR analysis of the corresponding LC fractions for the putative ligands revealed that 18β-glycyrrhetinic acid was the dominant isomer in the licorice extract (Supplemental Figure S2).


Novel Chemical Ligands to Ebola Virus and Marburg Virus Nucleoproteins Identified by Combining Affinity Mass Spectrometry and Metabolomics Approaches.

Fu X, Wang Z, Li L, Dong S, Li Z, Jiang Z, Wang Y, Shui W - Sci Rep (2016)

Structural elucidation of GC7 (A) and GC13 (B) by MS and MSMS analysis.The proposed fragmentation pathway is shown below mass spectra.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940736&req=5

f2: Structural elucidation of GC7 (A) and GC13 (B) by MS and MSMS analysis.The proposed fragmentation pathway is shown below mass spectra.
Mentions: Among the identified licorice constituents, we found GC7/10 (glycyrrhetinic acid) and GC13 (licochalcone A) met the aforementioned thresholds for both EBOV NP and MARV NP ligands (red symbols in Fig. 1C). It is noteworthy that GC7 (18β-glycyrrhetinic acid) and GC10 (18α-glycyrrhetinic acid) are a pair of diastereo-isomers differing only in their C18–H–, trans-, and cis-configuration, and they coeluted under the LC condition of this study. High-resolution MS and MSMS spectra were acquired on GC7/10 and GC13 in the protein incubation sample to support structural elucidation (Fig. 2). In addition, NMR analysis of the corresponding LC fractions for the putative ligands revealed that 18β-glycyrrhetinic acid was the dominant isomer in the licorice extract (Supplemental Figure S2).

Bottom Line: Accompanying biophysical analyses demonstrate that binding of 18β-glycyrrhetinic acid to EBOV NP significantly reduces protein thermal stability, induces formation of large NP oligomers, and disrupts the critical association of viral ssRNA with NP complexes whereas the compound showed no such activity on MARV NP.Our study has revealed the substantial potential of new analytical techniques in ligand discovery from natural herb resources.In addition, identification of a chemical ligand that influences the oligomeric state and RNA-binding function of EBOV NP sheds new light on antiviral drug development.

View Article: PubMed Central - PubMed

Affiliation: College of Biotechnology, Tianjin University of Science &Technology, Tianjin 300457, China.

ABSTRACT
The nucleoprotein (NP) of Ebola virus (EBOV) and Marburg virus (MARV) is an essential component of the viral ribonucleoprotein complex and significantly impacts replication and transcription of the viral RNA genome. Although NP is regarded as a promising antiviral druggable target, no chemical ligands have been reported to interact with EBOV NP or MARV NP. We identified two compounds from a traditional Chinese medicine Gancao (licorice root) that can bind both NPs by combining affinity mass spectrometry and metabolomics approaches. These two ligands, 18β-glycyrrhetinic acid and licochalcone A, were verified by defined compound mixture screens and further characterized with individual ligand binding assays. Accompanying biophysical analyses demonstrate that binding of 18β-glycyrrhetinic acid to EBOV NP significantly reduces protein thermal stability, induces formation of large NP oligomers, and disrupts the critical association of viral ssRNA with NP complexes whereas the compound showed no such activity on MARV NP. Our study has revealed the substantial potential of new analytical techniques in ligand discovery from natural herb resources. In addition, identification of a chemical ligand that influences the oligomeric state and RNA-binding function of EBOV NP sheds new light on antiviral drug development.

No MeSH data available.


Related in: MedlinePlus