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Assembly Pathway of Hepatitis B Core Virus-like Particles from Genetically Fused Dimers.

Holmes K, Shepherd DA, Ashcroft AE, Whelan M, Rowlands DJ, Stonehouse NJ - J. Biol. Chem. (2015)

Bottom Line: Macromolecular complexes are responsible for many key biological processes.However, in most cases details of the assembly/disassembly of such complexes are unknown at the molecular level, as the low abundance and transient nature of assembly intermediates make analysis challenging.Previous studies have proposed a trimer of dimers as an important intermediate species in assembly, acting to nucleate further assembly by dimer addition.

View Article: PubMed Central - PubMed

Affiliation: From the School of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom and.

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Related in: MedlinePlus

TEM of oligomer assembly reactions. Assembly was initiated by mixing samples of each complex with sodium chloride to a final concentration of 750 mm. Symbols 1, 2, and 3 relate to samples corresponding to fused Cp, and dimers or trimers of fused Cp, respectively. Micrographs (negative stain) of assembly reactions are shown, in the order corresponding to sample elution from size exclusion chromatography (see Fig. 3). The oligomers appear to be competent for assembly into capsids. Size bars, 100 nm.
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Figure 5: TEM of oligomer assembly reactions. Assembly was initiated by mixing samples of each complex with sodium chloride to a final concentration of 750 mm. Symbols 1, 2, and 3 relate to samples corresponding to fused Cp, and dimers or trimers of fused Cp, respectively. Micrographs (negative stain) of assembly reactions are shown, in the order corresponding to sample elution from size exclusion chromatography (see Fig. 3). The oligomers appear to be competent for assembly into capsids. Size bars, 100 nm.

Mentions: Assembly reactions, akin to those previously documented (7), were carried out to determine if the samples corresponding to dimers or trimers of fused Cp were competent for assembly into capsids. Samples of each protein at ∼0.2 mg ml−1 were treated with NaCl to a final concentration of 750 mm and assembly allowed to proceed for minimum of 2 h on ice. Protein samples were then analyzed by TEM which showed the presence of capsids in each of the samples (Fig. 5). This suggested that the three oligomers were competent for capsid assembly and, furthermore, that the oligomers were on pathway and were therefore assembly intermediates. However, it is possible that contamination of samples with very small amounts of free fused dimer Cp could be responsible for the results observed.


Assembly Pathway of Hepatitis B Core Virus-like Particles from Genetically Fused Dimers.

Holmes K, Shepherd DA, Ashcroft AE, Whelan M, Rowlands DJ, Stonehouse NJ - J. Biol. Chem. (2015)

TEM of oligomer assembly reactions. Assembly was initiated by mixing samples of each complex with sodium chloride to a final concentration of 750 mm. Symbols 1, 2, and 3 relate to samples corresponding to fused Cp, and dimers or trimers of fused Cp, respectively. Micrographs (negative stain) of assembly reactions are shown, in the order corresponding to sample elution from size exclusion chromatography (see Fig. 3). The oligomers appear to be competent for assembly into capsids. Size bars, 100 nm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4481223&req=5

Figure 5: TEM of oligomer assembly reactions. Assembly was initiated by mixing samples of each complex with sodium chloride to a final concentration of 750 mm. Symbols 1, 2, and 3 relate to samples corresponding to fused Cp, and dimers or trimers of fused Cp, respectively. Micrographs (negative stain) of assembly reactions are shown, in the order corresponding to sample elution from size exclusion chromatography (see Fig. 3). The oligomers appear to be competent for assembly into capsids. Size bars, 100 nm.
Mentions: Assembly reactions, akin to those previously documented (7), were carried out to determine if the samples corresponding to dimers or trimers of fused Cp were competent for assembly into capsids. Samples of each protein at ∼0.2 mg ml−1 were treated with NaCl to a final concentration of 750 mm and assembly allowed to proceed for minimum of 2 h on ice. Protein samples were then analyzed by TEM which showed the presence of capsids in each of the samples (Fig. 5). This suggested that the three oligomers were competent for capsid assembly and, furthermore, that the oligomers were on pathway and were therefore assembly intermediates. However, it is possible that contamination of samples with very small amounts of free fused dimer Cp could be responsible for the results observed.

Bottom Line: Macromolecular complexes are responsible for many key biological processes.However, in most cases details of the assembly/disassembly of such complexes are unknown at the molecular level, as the low abundance and transient nature of assembly intermediates make analysis challenging.Previous studies have proposed a trimer of dimers as an important intermediate species in assembly, acting to nucleate further assembly by dimer addition.

View Article: PubMed Central - PubMed

Affiliation: From the School of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom and.

Show MeSH
Related in: MedlinePlus