Three-dimensional structure of hepatitis B virus core particles determined by electron cryomicroscopy

Crowther, R. Anthony; Киселев, Н.А.; Böttcher, Bettina; Berriman, John; Borisova, Galina; Ose, Velta; Pumpens, Paul

doi:10.1016/0092-8674(94)90142-2

Cited by 479 publications

(450 citation statements)

References 23 publications

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“…Western blot analysis confirmed that eGFPs were successfully localized inside HBVC (Figure S2, Supporting Information). In wild‐type HBVC, two different types of icosahedral capsids have been observed: T = 3 (32 nm) and T = 4 (36 nm) symmetry‐based capsids consisting of 180 and 240 subunits, respectively 17. In this work, we used the truncated mutant (1–149 a.a.) of HBVC subunit, which reportedly prefers the T = 4 symmetry upon capsid formation 18.…”

Section: Resultsmentioning

confidence: 99%

Genetic Assembly of Double‐Layered Fluorescent Protein Nanoparticles for Cancer Targeting and Imaging

Kim

Kwon

et al. 2017

Advanced Science

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Hepatitis B virus capsid (HBVC), a self‐assembled protein nanoparticle comprised of 180 or 240 subunit proteins, is used as a cage for genetic encapsulation of fluorescent proteins (FPs). The self‐quenching of FPs is controlled by varying the spacing between FPs within the capsid structure. Double‐layered FP nanoparticle possessing cancer cell‐targeting capabilities is also produced by additionally attaching FPs and cancer cell receptor‐binding peptides (affibodies) to the outer surface of the capsid. The generically modified HBVC with double layers of mCardinal FPs and affibodies (mC‐DL‐HBVC) exhibit a high fluorescence intensity and a strong photostability, and is efficiently internalized by cancer cells and significantly stable against intracellular degradation. The mC‐DL‐HBVC effectively detects tumor in live mice with enhanced tumor targeting and imaging efficiency with far less accumulation in the liver, compared to a conventional fluorescent dye, Cy5.5. This suggests the great potential of mC‐DL‐HBVC as a promising contrast agent for in vivo tumor fluorescence imaging.

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Section: Resultsmentioning

confidence: 99%

Genetic Assembly of Double‐Layered Fluorescent Protein Nanoparticles for Cancer Targeting and Imaging

Kim

Kwon

et al. 2017

Advanced Science

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“…Moreover, no vesicle formation or plasma membrane invagination was detected by electron microscopy, confirming that the uptake is not mediated by endocytosis. Although the mechanism of translocation of cell-permeable peptides is controversial, 45 one model suggests that it occurs by formation of inverted micelles. 37 The structure labeled by a white arrow in the transmission electron microscopy image (Fig.…”

Section: Discussionmentioning

confidence: 99%

A novel system for efficient gene transfer into primary human hepatocytes via cell-permeable hepatitis B virus-like particle

et al. 2005

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R ecently, a variety of protein transduction domains (PTDs) have been identified allowing the transfer of peptides, proteins, or nucleic acids across cellular membranes into cells. 1-9 PTDs have been used to successfully treat preclinical models of human disease such as cancer, psoriasis, and stroke. 10-13 Transfer of nano-particular structures across cellular membranes is of increasing importance for the development of novel diagnostic and therapeutic tools. The capacity of PTDs to mediate an endocytosis-independent transfer of particles across the plasma membrane into the cytoplasm is still unclear. To investigate the potential of PTD to mediate the transfer of nano-particles across the cell membrane, virus-like particles (VLPs) harboring a marker gene were instrumental.Hepatitis B virus (HBV) core particles represent a very well-characterized VLP model system. 14,15 The HBV core particle (capsid) is assembled by 180 or 240 core protein monomers (HBcAg), resulting in an icosahedral particle 30 or 34 nm in diameter, respectively. A characteristic of the core protein is a basic arginine-rich C-terminal region, which is responsible for the packaging of DNA 16 and for guidance of the capsid to the nucleus. 17 Based on extensive structural analysis, 18 the hepatitis B capsid was used as a carrier for foreign epitopes to develop new vaccines. 19,20 In the viral context, the core particle harbors the viral genome (nucleocapsid). Efficient in vivo packaging of nucleic acids into capsids requires HBV polymerase and its interaction with a defined secondary structure (termed epsilon) at the 5Ј end of the RNA to be packaged. 21 The ⑀ signal is the primary element of the hepadnaviral packag-

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“…Cultivation of bacteria and purification of HBc⌬-derived proteins were based generally on the previously described method (11).…”

Section: Constructionmentioning

confidence: 99%

“…The high-resolution spatial structure of HBc icosahedrons (11,43) shows that the MIR is located on the tip of the spike, around the most protruding region between amino acids (aa) 78 and 82. For this reason, the MIR is generally accepted as the target site of choice for insertion of foreign epitopes (30).…”

mentioning

confidence: 99%

“…In many ways, HBc occupies a unique position among the VLP carriers because of its high-level synthesis and efficient selfassembly in virtually all known homologous and heterologous expression systems, including bacteria (for a review, see references 29 to 31). The major HBc B-cell epitopes (c and e1) are localized within the major immunodominant region (MIR), whereas the next important epitope, e2, is localized around amino acid position 130, close to the C-terminal histone-like region (for a review, see reference 30).The high-resolution spatial structure of HBc icosahedrons (11,43) shows that the MIR is located on the tip of the spike, around the most protruding region between amino acids (aa) 78 and 82. For this reason, the MIR is generally accepted as the target site of choice for insertion of foreign epitopes (30).…”

mentioning

confidence: 99%

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Construction and Immunological Evaluation of Multivalent Hepatitis B Virus (HBV) Core Virus-Like Particles Carrying HBV and HCV Epitopes

Sominskaya

Skrastiņa

Dislers

et al. 2010

Clin Vaccine Immunol

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A multivalent vaccine candidate against hepatitis B virus (HBV) and hepatitis C virus (HCV) infections was constructed on the basis of HBV core (HBc) virus-like particles (VLPs) as carriers. Chimeric VLPs that carried a virus-neutralizing HBV pre-S1 epitope corresponding to amino acids (aa) 20 to 47 in the major immunodominant region (MIR) and a highly conserved N-terminal HCV core epitope corresponding to aa 1 to 60 at the C terminus of the truncated HBc⌬ protein (N-terminal aa 1 to 144 of full-length HBc) were produced in Escherichia coli cells and examined for their antigenicity and immunogenicity. The presence of two different foreign epitopes within the HBc molecule did not interfere with its VLP-forming ability, with the HBV pre-S1 epitope exposed on the surface and the HCV core epitope buried within the VLPs. After immunization of BALB/c mice, specific T-cell activation by both foreign epitopes and a high-titer antibody response against the pre-S1 epitope were found, whereas an antibody response against the HBc carrier was notably suppressed. Both inserted epitopes also induced a specific cytotoxic-T-lymphocyte (CTL) response, as shown by the gamma interferon (IFN-␥) production profile.Genetically engineered virus-like particle (VLP)-based vaccines are one of the most promising tools in modern vaccinology. VLPs from almost all classes of viruses are being evaluated now or have just been adopted to use as carriers for presentation of foreign immunological epitopes (for a review, see references 29 and 31). VLP technologies possess obvious advantages for the generation of safe and efficacious vaccines. First, the repetitive antigenic structure of VLPs makes them highly immunogenic. Second, VLPs lack viral genomes or genes and are noninfectious, although they mimic infectious viruses in their structural and immunological features. Third, VLPs are generated by highly efficient heterologous expression of the cloned viral structural genes with subsequent quantitative in vivo or in vitro self-assembly of their products. Fourth, VLPs can be obtained by simple and efficient purification procedures. VLPs can be used for a broad range of applications, but the generation of vaccines against hepatitis B virus (HBV) and hepatitis C virus (HCV) infections is of special interest.The HBV core (HBc) protein was first reported as a promising VLP carrier in 1986 and was published in 1987 (6, 10, 24). In many ways, HBc occupies a unique position among the VLP carriers because of its high-level synthesis and efficient selfassembly in virtually all known homologous and heterologous expression systems, including bacteria (for a review, see references 29 to 31). The major HBc B-cell epitopes (c and e1) are localized within the major immunodominant region (MIR), whereas the next important epitope, e2, is localized around amino acid position 130, close to the C-terminal histone-like region (for a review, see reference 30).The high-resolution spatial structure of HBc icosahedrons (11,43) shows that the MIR is located on the tip of th...

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Three-dimensional structure of hepatitis B virus core particles determined by electron cryomicroscopy

Cited by 479 publications

References 23 publications

Genetic Assembly of Double‐Layered Fluorescent Protein Nanoparticles for Cancer Targeting and Imaging

Genetic Assembly of Double‐Layered Fluorescent Protein Nanoparticles for Cancer Targeting and Imaging

A novel system for efficient gene transfer into primary human hepatocytes via cell-permeable hepatitis B virus-like particle

Construction and Immunological Evaluation of Multivalent Hepatitis B Virus (HBV) Core Virus-Like Particles Carrying HBV and HCV Epitopes

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