Phosphotungstate as a useful eluent for hepatitis‐B virus surface antigen purification by heparin‐sepharose affinity chromatography

Tsutsumi, Yutaka; Nagumo, Fumio; Nishimura, Tadataka; Tadano, Jutaro

doi:10.1002/bmc.1130060206

Cited by 5 publications

(4 citation statements)

References 14 publications

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“…Use of lower-sulfated forms of heparin or CS resulted in either no inhibition (de-N-sulfated heparin) or dose-dependent reduction of HBV binding (CS and de-O-sulfated heparin). This is in line with previous publications using heparin-affinity chromatography for HBsAg purification (Einarsson et al, 1978;Tajima et al, 1992). Interestingly, binding of HBV to susceptible PTH and nonsusceptible PRH was only moderately inhibited by similar concentrations of highly sulfated polymers, such as heparin or dextran sulfate.…”

Section: Discussionsupporting

confidence: 91%

Role of glycosaminoglycans for binding and infection of hepatitis B virus

Leistner

Gruen-Bernhard

Glebe³

2007

Cell Microbiol

133

148

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SummaryMany parts of the life cycle of hepatitis B virus (HBV) infection of hepatocytes have been unravelled, but the attachment and entry process leading to infection is largely unknown. Using primary Tupaia hepatocyte cultures as an in vitro infection system, we determined that HBV uses cell-surface heparan sulfate proteoglycans as low-affinity receptor, because HBV infection was inhibited by heparin (IC50: 5 mg ml -1 ) or other higher-sulfated polymers, but not by lowersulfated glycosaminoglycans, such as chondroitin sulfate. Pretreatment of primary hepatocytes with heparinase decreased viral binding and inhibited HBV infection completely. Interestingly, after preS1-dependent viral binding at 16°C to the cell surface, subsequent infection could still be inhibited by HBV preS1-lipopeptides, but not by heparin any more, suggesting a shift of the virus to a high-affinity receptor. In summary, we suggest following multistep attachment process: in vivo, HBV is initially trapped within the liver in the space of Dissé by heparan sulfate proteoglycans. Thereafter, HBV binds via its preS1 attachment site and the N-terminal myristic acid to a yet unknown, high-affinity receptor that confers uptake in a yet unknown compartment.

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Section: Discussionsupporting

confidence: 91%

Role of glycosaminoglycans for binding and infection of hepatitis B virus

Leistner

Gruen-Bernhard

Glebe³

2007

Cell Microbiol

133

148

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“…5). HBsAg has previously been purified by heparin-affinity chromatography (Einarsson et al, 1978;Tajima et al, 1992) and we observed HBsAg in addition to intact virions in our heparin-purified HBV material (Fig. 4b).…”

Section: Discussionsupporting

confidence: 58%

Hepatitis C virus and hepatitis B virus bind to heparin: purification of largely IgG-free virions from infected plasma by heparin chromatography

Zahn

Allain

2005

Journal of General Virology

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Binding to heparin of hepatitis C virus (HCV) and hepatitis B virus (HBV) from chronic carriers was investigated. Eighty per cent of HCV RNA from an agammaglobulinaemic patient (IgG-free virus) was retained on immobilized heparin and eluted with ¢0?4 M NaCl, in contrast tõ 20 % from immunocompetent chronic carriers (with ¡8 % IgG-free virus). Increased binding to heparin of the HCV fraction that was not retained by a protein G column suggested that antibodies complexed to the virions partially inhibited the interaction. A higher proportion (15-80 %) of HBV from chronic carriers bound to heparin and eluted with ¢0?4 M NaCl. After washing of the heparin columns with 0?3 M NaCl, <1 % of total plasma proteins co-eluted with HCV or HBV. By this one-step heparin chromatography, without ultracentrifugation, IgG-free HCV and IgG-free HBV were preferentially purified from human plasma by 1000-fold and greater than 500-fold, respectively. Following assessment with an anti-E2 envelope protein antibody, the amount of immunoprecipitated HCV particles after heparin purification was similar to that in the original plasma, suggesting that undamaged virions were purified. This was further supported by heparin-purified HCV binding to lymphocyte cell lines in a dose-dependent manner. Intact HBV particles were detected by electron microscopy. It was concluded that HCV and HBV from chronically infected patients bind to heparin, the closest homologue of liver heparan sulfate, and that heparin chromatography is an efficient and gentle method for purifying these viruses from human plasma. In the absence of cell-culture systems or alternative robust purification methods, heparin chromatography may help greatly in binding and infectivity studies.

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“…The high anionic valence present in such a small area is responsible for the strong charge interaction between PW 11 and its target molecule(s), which is hardly affected by steric hindrance. For instance, our previous studies demonstrated that the saltingin effect and elution power in affinity chromatography of PW 11 were 100~1000 fold higher than those of NaCl [38][39][40]. It is now considered that such compounds may be useful as anionic probes, which will affect certain biochemical and/or biological systems where charge interaction plays an important role [41].…”

Section: The History Of β-Lactam Resistance In Staphylococcus Aureusmentioning

confidence: 99%

“…POTs will interfere with certain binding systems where charge interaction plays an important role [41]. For instance, these compounds may be useful as eluents for affinity chromatography [39,40]. It is reported that SiW 11 /SiW 12 efficiently solubilizes some components from subcellular organelles [136].…”

Section: Acmentioning

confidence: 99%

Polyoxotungstates Reduce the β-Lactam Resistance of Methicillin-Resistant Staphylococcus aureus

Tsutsumi

2005

MRMC

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Bacterial strains isolated from clinical specimens have become more and more resistant to many anti-microbials. This is because we have consumed large amounts of strong antimicrobials over long periods of time and thus bacterial cells are able to survive by altering the target(s) of antimicrobial agents. A good example of this phenomenon is methicillin-resistant Staphylococcus aureus (MRSA). One of the cell wall-synthesizing enzymes (known as PBP2') of this pathogen has low affinity to beta-lactams, and therefore the bacterial cells continue to grow even under high concentrations of the agents. However, this drug resistance does not seem to be total. They seem to have some weak spots and several substances are known to sensitize strains of MRSA to beta-lactams. This review discusses the ability of polyoxotungstates (POTs) to sensitize MRSA to beta-lactams by reducing the expression of PBP2'. It is also possible that the sensitization is a type of stress response of MRSA to POTs. This idea may provide a hint for the development of a new antimicrobial agent.

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Phosphotungstate as a useful eluent for hepatitis‐B virus surface antigen purification by heparin‐sepharose affinity chromatography

Cited by 5 publications

References 14 publications

Role of glycosaminoglycans for binding and infection of hepatitis B virus

Role of glycosaminoglycans for binding and infection of hepatitis B virus

Hepatitis C virus and hepatitis B virus bind to heparin: purification of largely IgG-free virions from infected plasma by heparin chromatography

Polyoxotungstates Reduce the β-Lactam Resistance of Methicillin-Resistant Staphylococcus aureus

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Phosphotungstate as a useful eluent for hepatitis‐B virus surface antigen purification by heparin‐sepharose affinity chromatography

Cited by 5 publications

References 14 publications

Role of glycosaminoglycans for binding and infection of hepatitis B virus

Role of glycosaminoglycans for binding and infection of hepatitis B virus

Hepatitis C virus and hepatitis B virus bind to heparin: purification of largely IgG-free virions from infected plasma by heparin chromatography

Polyoxotungstates Reduce the &#946;-Lactam Resistance of Methicillin-Resistant Staphylococcus aureus

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Polyoxotungstates Reduce the β-Lactam Resistance of Methicillin-Resistant Staphylococcus aureus