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An N-terminal extension to the hepatitis B virus core protein forms a poorly ordered trimeric spike in assembled virus-like particles.

McGonigle R, Yap WB, Ong ST, Gatherer D, Bakker SE, Tan WS, Bhella D - J. Struct. Biol. (2014)

Bottom Line: Heterologous sequences have been successfully inserted at both amino and carboxy termini as well as internally at the major immunodominant epitope.The insert was seen to form a trimeric spike on the capsid surface that was poorly resolved, most likely owing to it being flexible.We hypothesise that the capacity of N-terminal inserts to form trimers may have application in the development of multivalent vaccines to trimeric antigens.

View Article: PubMed Central - PubMed

Affiliation: MRC-University of Glasgow Centre for Virus Research, Sir Michael Stoker Building, Garscube Campus, 464 Bearsden Road, Glasgow G61 1QH, Scotland, UK.

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Related in: MedlinePlus

Blocres local resolution analysis of His-β-L HBcAg VLP reconstructions. Local FSC resolution analysis was performed using the BSOFT routine Blocres. The T = 3 (A) and T = 4 (B) reconstructions are presented in stereo pair view and coloured according to local resolution. For both structures, the N-terminal inserted region, has a lower resolution than the native HBcAg capsid. This finding is consistent with the ResMap analysis and suggests that the inserted region is more flexible than the core capsid structure. Unlike the ResMap analyses however the HBcAg dimeric spikes are assessed as being higher-resolution although the distal portions of these four-helix bundles are lower resolution than the shell of the capsid (C and D).
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f0015: Blocres local resolution analysis of His-β-L HBcAg VLP reconstructions. Local FSC resolution analysis was performed using the BSOFT routine Blocres. The T = 3 (A) and T = 4 (B) reconstructions are presented in stereo pair view and coloured according to local resolution. For both structures, the N-terminal inserted region, has a lower resolution than the native HBcAg capsid. This finding is consistent with the ResMap analysis and suggests that the inserted region is more flexible than the core capsid structure. Unlike the ResMap analyses however the HBcAg dimeric spikes are assessed as being higher-resolution although the distal portions of these four-helix bundles are lower resolution than the shell of the capsid (C and D).

Mentions: To confirm our findings, we evaluated local resolution within the maps using an alternative method: the Bsoft routine Blocres (Cardone et al., 2013). This approach calculates Fourier shell correlation values for a sliding box to yield a resolution map (Fig. 3). Supporting the results obtained with ResMap, Blocres determined that the T = 3 structure was resolved at between 6 and 9 Å within the HBcAg region while the N-terminal extension was estimated to be resolved at between 10 and 11 Å. The resolution estimates for the T = 4 VLP were also consistent with the ResMap output, measuring between 8 and 10 Å within the core components while the structure of the putative trimeric spike was solved at poorer than 14 Å resolution.


An N-terminal extension to the hepatitis B virus core protein forms a poorly ordered trimeric spike in assembled virus-like particles.

McGonigle R, Yap WB, Ong ST, Gatherer D, Bakker SE, Tan WS, Bhella D - J. Struct. Biol. (2014)

Blocres local resolution analysis of His-β-L HBcAg VLP reconstructions. Local FSC resolution analysis was performed using the BSOFT routine Blocres. The T = 3 (A) and T = 4 (B) reconstructions are presented in stereo pair view and coloured according to local resolution. For both structures, the N-terminal inserted region, has a lower resolution than the native HBcAg capsid. This finding is consistent with the ResMap analysis and suggests that the inserted region is more flexible than the core capsid structure. Unlike the ResMap analyses however the HBcAg dimeric spikes are assessed as being higher-resolution although the distal portions of these four-helix bundles are lower resolution than the shell of the capsid (C and D).
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4318616&req=5

f0015: Blocres local resolution analysis of His-β-L HBcAg VLP reconstructions. Local FSC resolution analysis was performed using the BSOFT routine Blocres. The T = 3 (A) and T = 4 (B) reconstructions are presented in stereo pair view and coloured according to local resolution. For both structures, the N-terminal inserted region, has a lower resolution than the native HBcAg capsid. This finding is consistent with the ResMap analysis and suggests that the inserted region is more flexible than the core capsid structure. Unlike the ResMap analyses however the HBcAg dimeric spikes are assessed as being higher-resolution although the distal portions of these four-helix bundles are lower resolution than the shell of the capsid (C and D).
Mentions: To confirm our findings, we evaluated local resolution within the maps using an alternative method: the Bsoft routine Blocres (Cardone et al., 2013). This approach calculates Fourier shell correlation values for a sliding box to yield a resolution map (Fig. 3). Supporting the results obtained with ResMap, Blocres determined that the T = 3 structure was resolved at between 6 and 9 Å within the HBcAg region while the N-terminal extension was estimated to be resolved at between 10 and 11 Å. The resolution estimates for the T = 4 VLP were also consistent with the ResMap output, measuring between 8 and 10 Å within the core components while the structure of the putative trimeric spike was solved at poorer than 14 Å resolution.

Bottom Line: Heterologous sequences have been successfully inserted at both amino and carboxy termini as well as internally at the major immunodominant epitope.The insert was seen to form a trimeric spike on the capsid surface that was poorly resolved, most likely owing to it being flexible.We hypothesise that the capacity of N-terminal inserts to form trimers may have application in the development of multivalent vaccines to trimeric antigens.

View Article: PubMed Central - PubMed

Affiliation: MRC-University of Glasgow Centre for Virus Research, Sir Michael Stoker Building, Garscube Campus, 464 Bearsden Road, Glasgow G61 1QH, Scotland, UK.

Show MeSH
Related in: MedlinePlus