Phosphotungstate as a useful eluent for antithrombin III purification by heparin-agarose affinity chromatography

Tsutsumi, Yutaka; Shizuka, Reiko; Oshitani, Setsuko; Amagai, H

doi:10.1016/0049-3848(90)90027-a

Cited by 7 publications

(9 citation statements)

References 13 publications

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“…lb). A similar phenomenon was also observed in the purification of ATTTI (Tajima et al, 1990). In order to elute the residual proteins remaining in the column, a NaCl solution with a high ionic strength (over 500 mM) was required.…”

Section: Purification Procedures (Procedures A)mentioning

confidence: 55%

“…The molecular weight markers were products of Bio-Rdd (Richmond, VA, USA). The gel was stained with silver by a modification of Morrissey's method (Morrissey, 1981), as described in our previous report (Tajima et al, 1990).…”

Section: Resultsmentioning

confidence: 99%

“…However, PTA has a heparin-like effect and acts potently on the anion(heparin)-binding site of AT111 at lower concentrations than NaC1. It seems that PTA reduces the affinity of AT111 for heparin as well as simply competing with heparin for the binding site (Tajima et al, 1989(Tajima et al, , 1990. We considered therefore that PTA might be useful for the purification of HBsAg and consequently carried out of study of the effect of PTA elution for HBsAg adsorbed on affinity gel.…”

Section: Introduction Experimentalmentioning

confidence: 97%

“…We have previously reported that PTA is more specific than NaCl for the purification of antithrombin TI1 (ATITI) using heparin-Sepharose (Tajima et al, 1990). The intraction between ATIII and heparin on the gel is very strong.…”

Section: Introduction Experimentalmentioning

confidence: 98%

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

Tsutsumi

Nagumo

Nishimura

et al. 1992

Biomedical Chromatography

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On the basis of the heparin-like effect of phosphotungstate (PTA), we have shown that it is useful for the purification of hepatitis-B virus surface antigen (HBsAg) using heparin-Sepharose affinity chromatography. HBsAg was eluted with 0.2-0.6 M NaCl. HBsAg was also eluted with PTA at approximately 1 mM, and the HBsAg fraction thus obtained contained fewer impurities than the corresponding fraction eluted with NaCl. Moreover, PTA yielded HBsAg and hepatitis-B virus e-antigen simultaneously. PTA may specifically reduce the affinity of HBsAg for heparin as well as simply competing with heparin for an anion-binding site of HBsAg. Residual PTA in the eluate was easily decomposed by alkalization, which was useful for subsequent studies.

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Section: Purification Procedures (Procedures A)mentioning

confidence: 55%

Section: Resultsmentioning

confidence: 99%

Section: Introduction Experimentalmentioning

confidence: 97%

Section: Introduction Experimentalmentioning

confidence: 98%

See 2 more Smart Citations

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

Tsutsumi

Nagumo

Nishimura

et al. 1992

Biomedical Chromatography

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“…Oni poseduju antibakterijska (5, 6), antiviralna (7-11), antitumorska (12) i antikoagulantna (13,14) svojstva. Među njihovim svojstvima izdvajaju se potentnost i visoka selektivnost kao inhibitora acetilholinesterase (15).…”

Section: Uvodunclassified

modulation of activity of acetylcholinesterase with polyoxovolframs

Lacković¹,

Stojicic²,

Lalatovic³

et al. 2017

Med podmladak

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Introduction: Alzheimer's disease has its pathoanatomical origin. The loss of synapses and neurons is dominant in this disease and it leads to changes, primarily at the acetylcholine neurotransmitter system. Polyoxometalates (POM) are highly potent and selective inhibitors of acetylcholinesterase, the enzyme responsible for the disintegration of acetylcholine, and therefore they find their usage in the treatment of Alzheimer's disease, since it would increase and extend the effect of the missing acetylcholine. The aim: Presenting the results of in vitro influence of POM on the activity of AChE. Material and methods:In our study we have tested the effects of five Heteropolyoxovolframs: Results:The tested compounds make the inhibition of AChE activity in a dose-dependent manner. Inhibition parameters obtained in this study demonstrate that the compounds 2, 3 and 5 are very potent inhibitors of the enzyme activity. On the other hand, compounds 1 and 4 demonstrate about a hundred, or a thousand times, less potent inhibitory effect on the activity of AchE, compering to the most potent inhibitor (compound 3), whose IC50 value is 4,79×10 -7 mol/L. The results obtained in this study are consistent with previously demonstrated inhibitory effect of polyoxovolframs on cholinesterase activity. Conclusion: According to the obtained results we can conclude that the tested Polyoxometalates at physiological pH values inhibit the activity of AChE, and that the inhibition is dose-dependent. Inhibitory range of tested POM in this study verify significantly, possibly as a result of differences in charge, size and shape of these POM.

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Transforming growth factor‐β1 is a heparin‐binding protein: Identification of putative heparin‐binding regions and isolation of heparins with varying affinity for TGF‐β1

McCaffrey

Falcone

1992

Journal Cellular Physiology

176

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Previous studies indicated that a major factor in heparin's ability to suppress the proliferation of vascular smooth muscle cells is an interaction with transforming growth factor-beta 1 (TGF-beta 1). Heparin appeared to bind directly to TGF-beta 1 and to prevent the association of TGF-beta 1 with alpha 2-macroglobulin (alpha 2-M). The present studies indicate that 20-70% of iodinated TGF-beta 1 binds to heparin-Sepharose and the retained fraction is eluted with approximately 0.37 M NaCl. Native, unlabelled platelet TGF-beta 1, however, is completely retained by heparin-Sepharose and eluted with 0.9-1.2 M NaCl. Using synthetic peptides, the regions of TGF-beta 1 that might be involved in the binding of heparin and other polyanions were examined. Sequence analysis of TGF-beta 1 indicated three regions with a high concentration of basic residues. Two of these regions had the basic residues arranged in a pattern homologous to reported consensus heparin-binding regions of other proteins. The third constituted a structurally novel pattern of basic residues. Synthetic peptides homologous to these three regions, but not to other regions of TGF-beta 1, were found to bind to heparin-Sepharose and were eluted with 0.15 M-0.30 M NaCl. Only two of these regions were capable of blocking the binding of heparin to 125I-TGF-beta. Immobilization of these peptides, followed by affinity purification of heparin, indicated that one peptide was capable of isolating subspecies of heparin with high and low affinity for authentic TGF-beta 1. The ability of TGF-beta 1 to bind to heparin or related proteoglycans under physiological conditions may be useful in understanding the biology of this pluripotent growth and metabolic signal. Conversely, a subspecies of heparin molecules with high affinity for TGF-beta 1 may be a factor in some of the diverse biological actions of heparin.

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Phosphotungstate as a useful eluent for antithrombin III purification by heparin-agarose affinity chromatography

Cited by 7 publications

References 13 publications

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

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

modulation of activity of acetylcholinesterase with polyoxovolframs

Transforming growth factor‐β1 is a heparin‐binding protein: Identification of putative heparin‐binding regions and isolation of heparins with varying affinity for TGF‐β1

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