Failure to detect polyalbumin-binding sites on the woodchuck hepatitis virus surface antigen: implications for the pathogenesis of hepatitis B virus in humans

Pohl, Christoph; Côté, Paul J.; Purcell, Robert H.; Gerin, John L.

doi:10.1128/jvi.60.3.943-949.1986

Cited by 14 publications

(4 citation statements)

References 35 publications

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“…It appears a worthwhile goal to search for cell-culture systems able to bind pHSA or to isolate the pHSA receptors from natural human liver. The hepadnavirus of the woodchuck, however, does not bind natural or polymerized woodchuck serum albumin (39). This shows that albumin binding cannot be a general mechanism of the hepadnaviral life cycle; it prevents use of animal hepadnaviruses for the further study of this phenomenon.…”

Section: Discussionmentioning

confidence: 96%

Interaction between hepatitis B surface proteins and monomeric human serum albumin

et al. 1990

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HBsAg is known to bind to human serum albumin polymerized by glutaraldehyde, human serum albumin has been found in preparations of HBsAg by several investigators. However, it is not yet known whether natural human serum albumin binds to hepatitis B virus under physiological conditions. We studied the binding between natural or recombinant HBsAg and monomeric human serum albumin by immunological, biochemical and biophysical methods. The binding capacity of 20-nm HBs spheres was variable but ranged up to six molecules HSA/sphere. A reversible binding site for human serum albumin was exclusively localized in the preS2 domain, whereas the S domain was inactive in vitro. Human serum albumin copurified with HBsAg of human origin during gel chromatography or sucrose-gradient centrifugation. This human serum albumin was monomeric in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The preS2-bound part of the human serum albumin could be removed from HBsAg by high-salt, such as CsCl centrifugation, but another part could only be removed by treatment with a disulfide cleaving reagent. Most of this covalently bound human serum albumin was retained at the HBsAg particle after complete cleavage of medium-sized HBs protein with trypsin. This indicates a second way in which albumin binds irreversible to cysteine(s) of the small HBs protein (SHBs, P24 and GP27).

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Section: Discussionmentioning

confidence: 96%

Interaction between hepatitis B surface proteins and monomeric human serum albumin

et al. 1990

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“…Feitelson et al suggested that besides the major protein bands of SWHs protein, larger proteins with the pre-S sequences were present, but a clear identification was not possible (10). Pohl et al reported that the protein composition of WHsAg from serum was quite similar to those of the pre-S-containing proteins gp33, gp36, p39, and gp42 of HBsAg, but actually the WHs protein band in their silverstained SDS gel corresponding to gp33 of HBsAg was very weak, and the other proteins were not clearly resolved and possibly larger (30). Schaeffer et al identified by immunoblotting with an antiserum against WHV pre-S peptide (residues 85 to 173) four proteins in partially purified WHsAg of 33, 36, 45, and 47 kDa.…”

Section: Discussionmentioning

confidence: 99%

“…(ii) The human enhancer I of HBV is obviously not present in the WHV genome (9). (iii) MWHs protein does not bind woodchuck serum albumin (30,34), whereas MHBs protein binds in a species-specific manner modified natural human serum albumin (21). (iv) The LWHs protein is 42 (or 31) amino acids longer than the LHBs protein, due to a larger pre-S1 domain.…”

Section: Discussionmentioning

confidence: 99%

“…The protein composition of WHsAg has been studied previously (10,30,33,35), but the identification of the larger protein bands was incomplete. Since the unexplained protein pattern of WHsAg in SDS gel electrophoresis led to the question of whether WHV has a completely different expression strategy for its surface proteins, we determined their primary structure and searched for posttranslational modifications different from those in HBs proteins.…”

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Structure and Glycosylation Patterns of Surface Proteins from Woodchuck Hepatitis Virus

Tolle

Glebe

Linder

et al. 1998

J Virol

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Woodchucks chronically infected with woodchuck hepatitis virus (WHV) are a valuable model for human hepatitis B virus (HBV) in studies of pathogenesis, immunity, and antiviral therapy. For this reason, substantial efforts to characterize both the similarities and the differences between HBV and WHV are being made. The structure of the WHV surface proteins (WHs proteins) has not yet been adequately elucidated. The bands that would be expected for glycosylated and nonglycosylated small (S) WHs protein are found by sodium dodecyl sulfate gel electrophoresis of purified WHs protein, but the bands corresponding to the middle (M) and large (L) WHs proteins of HBV are not seen at the expected sizes, even though the sequences of the WHV and HBV surface protein genes are 60% homologous. By amino-terminal sequencing we have identified two bands at 41 and 45 kDa as the MWHs proteins, 8 kDa larger than expected. We have also confirmed that two bands at 24 and 27 kDa are SWHs proteins. A protein of 49 kDa was blocked at the N terminus, which using immunoblotting with an antiserum against WHV pre-S1 (positions 126 to 146) was identified, together with a part of the 45-kDa protein, as glycosylated and nonglycosylated LWHs protein of the expected size. Sialidase and O-glycosidase digestion showed that the larger size of MWHs protein results from the presence of O glycoside groups which are probably in the pre-S2 domain of MWHs protein. Since the pre-S2 domains of HBV and WHV have similar numbers of potential O glycosylation sites, it appears to be likely that the glycosyltransferases act differently on the viral proteins in woodchucks and humans.

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α-fetoprotein in the woodchuck model of hepadnavirus infection and disease: Immunochemical analysis of woodchuck α-fetoprotein and measurement in serum by quantitative monoclonal radioimmunoassay

et al. 1990

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Woodchuck hepatitis virus infection of the eastern woodchuck represents a useful model for the study of hepatitis B virus infection and disease in humans, including hepatocellular carcinoma. In man, hepatocellular carcinoma is frequently detected and monitored using assays for serum alpha-fetoprotein. To study the relationship between alpha-fetoprotein and woodchuck hepatitis virus-induced hepatocellular carcinoma in the woodchuck model, we produced a monoclonal antibody to woodchuck alpha-fetoprotein and used biophysical and immunochemical methods to demonstrate its specificity and affinity (7 x 10(8) L/mol) for woodchuck alpha-fetoprotein. A competition radioimmunoassay was then developed and standardized for measuring serum alpha-fetoprotein concentrations. In the radioimmunoassay system, woodchuck alpha-fetoprotein was detected between 20 ng/ml (20% to 25% inhibition) and 8,500 ng/ml (97% to 98% inhibition). Elevated serum alpha-fetoprotein concentrations (450 to 452,000 ng/ml) were measured in 21 of 23 woodchucks in the advanced stages of woodchuck hepatitis virus-induced hepatocellular carcinoma. Serum alpha-fetoprotein was elevated above normal (greater than or equal to 450 ng/ml) as early as 3 to 11 mo before terminal hepatocellular carcinoma in 11 of 16 of the woodchuck hepatitis virus-carrier woodchucks. In a pilot study, serum alpha-fetoprotein became markedly elevated above normal in woodchuck hepatitis virus-carrier woodchucks that developed hepatocellular carcinoma but not in serologically recovered or uninfected woodchucks (i.e., without hepatocellular carcinoma). Thus, alpha-fetoprotein may provide a useful noninvasive marker in the woodchuck model for detecting and monitoring woodchuck hepatitis virus-induced hepatocellular carcinoma from earlier stages.

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Failure to detect polyalbumin-binding sites on the woodchuck hepatitis virus surface antigen: implications for the pathogenesis of hepatitis B virus in humans

Cited by 14 publications

References 35 publications

Interaction between hepatitis B surface proteins and monomeric human serum albumin

Interaction between hepatitis B surface proteins and monomeric human serum albumin

Structure and Glycosylation Patterns of Surface Proteins from Woodchuck Hepatitis Virus

α-fetoprotein in the woodchuck model of hepadnavirus infection and disease: Immunochemical analysis of woodchuck α-fetoprotein and measurement in serum by quantitative monoclonal radioimmunoassay

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