Limits...
A C-Terminal Hydrophobic Region is Required for Homo-Oligomerization of the Hepatitis E Virus Capsid (ORF2) Protein.

Xiaofang L, Zafrullah M, Ahmad F, Jameel S - J. Biomed. Biotechnol. (2001)

Bottom Line: When expressed in vitro or in transfected cells, the ORF2 protein assembled as dimers, trimers and higher order forms.While N-terminal deletions upto 111 amino acids had no effect, the deletion of amino acids 585-610 led to reduced homo-oligomerization.This deletion also resulted in aberrant folding of the protein, as determined by its sensitivity to trypsin.This study suggests that a C-terminal hydrophobic region encompassing amino acids 585-610 of the ORF2 protein might be critical for capsid biogenesis.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
Hepatitis E virus (HEV) is the causative agent of hepatitis E, an acute form of viral hepatitis. The open reading frame 2 (ORF2) of HEV encodes the viral capsid protein, which can self-oligomerize into virus-like particles. To understand the domains within this protein important for capsid biogenesis, we have carried out in vitro analyses of association and folding patterns of wild type and mutant ORF2 proteins. When expressed in vitro or in transfected cells, the ORF2 protein assembled as dimers, trimers and higher order forms.While N-terminal deletions upto 111 amino acids had no effect, the deletion of amino acids 585-610 led to reduced homo-oligomerization. This deletion also resulted in aberrant folding of the protein, as determined by its sensitivity to trypsin. This study suggests that a C-terminal hydrophobic region encompassing amino acids 585-610 of the ORF2 protein might be critical for capsid biogenesis.

No MeSH data available.


Related in: MedlinePlus

The 660 amino acid protein (pORF2) encoded by the HEV ORF2 is shown schematically, with three major regions: the extreme N-terminus, containing the putative signal sequence (open box), the highly basic N-terminal half of the protein (+) and a C-terminal hydrophobic stretch (filled box). The positions of three potential N-linked glycosylation sites are shown at amino acid residues 137, 310, 562. The deletion mutants used inthis study are also shown schematically.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC129057&req=5

Figure 1: The 660 amino acid protein (pORF2) encoded by the HEV ORF2 is shown schematically, with three major regions: the extreme N-terminus, containing the putative signal sequence (open box), the highly basic N-terminal half of the protein (+) and a C-terminal hydrophobic stretch (filled box). The positions of three potential N-linked glycosylation sites are shown at amino acid residues 137, 310, 562. The deletion mutants used inthis study are also shown schematically.

Mentions: The expression vectors pSG-ORF2, pSG-ORF2[Δ2–34]and pSG-ORF2[137/310/562], expressing the wild type, signal sequence-deleted and glycosylation- ORF2 proteins, respectively, have been described earlier [9]. The ORF2[Δ2–111] mutant was generated by digesting ORF2 at a SalI site (nucloetide 381), followed by oligonucleotide-based reconstruction. The expression vectors pSG-ORF2[Δ585–610] and pSG-ORF2[Δ2–111/Δ585–610] were generated by deleting a C-terminal hydrophobic region encompassing amino acids 585–610 by site-directed mutgenesis Figure 1.


A C-Terminal Hydrophobic Region is Required for Homo-Oligomerization of the Hepatitis E Virus Capsid (ORF2) Protein.

Xiaofang L, Zafrullah M, Ahmad F, Jameel S - J. Biomed. Biotechnol. (2001)

The 660 amino acid protein (pORF2) encoded by the HEV ORF2 is shown schematically, with three major regions: the extreme N-terminus, containing the putative signal sequence (open box), the highly basic N-terminal half of the protein (+) and a C-terminal hydrophobic stretch (filled box). The positions of three potential N-linked glycosylation sites are shown at amino acid residues 137, 310, 562. The deletion mutants used inthis study are also shown schematically.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC129057&req=5

Figure 1: The 660 amino acid protein (pORF2) encoded by the HEV ORF2 is shown schematically, with three major regions: the extreme N-terminus, containing the putative signal sequence (open box), the highly basic N-terminal half of the protein (+) and a C-terminal hydrophobic stretch (filled box). The positions of three potential N-linked glycosylation sites are shown at amino acid residues 137, 310, 562. The deletion mutants used inthis study are also shown schematically.
Mentions: The expression vectors pSG-ORF2, pSG-ORF2[Δ2–34]and pSG-ORF2[137/310/562], expressing the wild type, signal sequence-deleted and glycosylation- ORF2 proteins, respectively, have been described earlier [9]. The ORF2[Δ2–111] mutant was generated by digesting ORF2 at a SalI site (nucloetide 381), followed by oligonucleotide-based reconstruction. The expression vectors pSG-ORF2[Δ585–610] and pSG-ORF2[Δ2–111/Δ585–610] were generated by deleting a C-terminal hydrophobic region encompassing amino acids 585–610 by site-directed mutgenesis Figure 1.

Bottom Line: When expressed in vitro or in transfected cells, the ORF2 protein assembled as dimers, trimers and higher order forms.While N-terminal deletions upto 111 amino acids had no effect, the deletion of amino acids 585-610 led to reduced homo-oligomerization.This deletion also resulted in aberrant folding of the protein, as determined by its sensitivity to trypsin.This study suggests that a C-terminal hydrophobic region encompassing amino acids 585-610 of the ORF2 protein might be critical for capsid biogenesis.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
Hepatitis E virus (HEV) is the causative agent of hepatitis E, an acute form of viral hepatitis. The open reading frame 2 (ORF2) of HEV encodes the viral capsid protein, which can self-oligomerize into virus-like particles. To understand the domains within this protein important for capsid biogenesis, we have carried out in vitro analyses of association and folding patterns of wild type and mutant ORF2 proteins. When expressed in vitro or in transfected cells, the ORF2 protein assembled as dimers, trimers and higher order forms.While N-terminal deletions upto 111 amino acids had no effect, the deletion of amino acids 585-610 led to reduced homo-oligomerization. This deletion also resulted in aberrant folding of the protein, as determined by its sensitivity to trypsin. This study suggests that a C-terminal hydrophobic region encompassing amino acids 585-610 of the ORF2 protein might be critical for capsid biogenesis.

No MeSH data available.


Related in: MedlinePlus