Use of exoglycosidases from Mercenaria mercenaria (hard shelled clam) as a tool for structural studies of glycosphingolipids and glycoproteins

Ghosh, Sujoy; Lee, Sunmin; Brown, Thomas A.; Basu, Manju; Hawes, John W.; Davidson, Donald J.; Basu, Subhash

doi:10.1016/0003-2697(91)90462-3

Cited by 11 publications

(4 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An equal volume of 5 % (w\v) trichloroacetic acid was added, and the mixture was left at room temperature for 10 min and then centrifuged at 16 000 g for 3 min. Aliquots of the supernatant were removed and mixed with an equal volume of 0.5 M NaOH, and the absorbance was read at 405 nm [9]. The percentage inhibition was calculated from a standard curve of trypsin activity done in parallel.…”

Section: Inhibition Of Trypsin Activitymentioning

confidence: 99%

Modification of the erythrocyte surface in rats bearing Yoshida ascites sarcoma is brought about by a tumour variant of α2-macroglobulin

Sanjay

Kalraiya

Mehta

1997

Biochemical Journal

View full text Add to dashboard Cite

Erythrocytes from the circulation of rats bearing Yoshida ascites sarcoma exhibit higher concanavalin A (ConA)-mediated agglutinability than those from normal animals. A tetrameric glycoprotein of subunit molecular mass 170 kDa, purified from the cell-free ascites fluid, was found to confer higher ConA-mediated agglutinability on erythrocytes in vitro. An antiserum to this tumour-derived protein failed to detect any cross-reactive component in normal rat plasma or in any of the normal tissues examined. An immunoreactive protein was, however, detected in blood plasma when the acute-phase reaction was stimulated by injection of turpentine. The cross-reactive acute-phase protein was purified by ConA-affinity, gel-filtration and ion-exchange chromatography, and identified as alpha2-macroglobulin. The acute-phase protein and the protein obtained from the ascites fluid have identical or very similar native and subunit molecular masses, subunit arrangement and pI. They both are able to inhibit trypsin and, as a consequence, acquire greater mobility in native PAGE. In addition, the two proteins bind to rat erythrocytes non-specifically, and in similar amounts. However, despite these similarities, the acute-phase protein is unable to enhance the agglutinability of erythrocytes. The two proteins differ in their carbohydrate content, but this differential glycosylation is not the cause of the difference in their surface modification activity. The chemically deglycosylated proteins show a small but consistent difference in the size of their polypeptides. Their tryptic peptide maps, although largely similar, show some differences, as do their amino acid compositions. It is probable that the proteins are independent members of the same (alpha-macroglobulin) family. The rat embryo is also found to express a soluble protein consisting of a 170 kDa polypeptide that cross-reacts with the antibody to the tumour-derived protein. The purified embryo protein is able to alter the ConA-mediated agglutinability of erythrocytes in vitro, and also yields a tryptic peptide map that is identical to that of the tumour-derived protein. The modification of the host cell surface in the tumour-bearing rats is thus caused by what appears to be a tumour (oncofetal?) variant of alpha2-macroglobulin.

show abstract

Section: Inhibition Of Trypsin Activitymentioning

confidence: 99%

Modification of the erythrocyte surface in rats bearing Yoshida ascites sarcoma is brought about by a tumour variant of α2-macroglobulin

Sanjay

Kalraiya

Mehta

1997

Biochemical Journal

View full text Add to dashboard Cite

show abstract

“…The clam glycosidase mixture, used for total enzymatic digestions ofasialyl-oligosaccharides and for removal ofO-linked oligosaccharide from AN-HPg or AN-ir-HPg, was prepared as described previously ( 15). The fraction used in this investigation was that from the DEAE CL-6B Sepharose anion-exchange chromatography column, which contained at least 16 different glycosidases (15).…”

Section: Introductionmentioning

confidence: 99%

“…The fraction used in this investigation was that from the DEAE CL-6B Sepharose anion-exchange chromatography column, which contained at least 16 different glycosidases (15). This mixture was utilized under conditions previously detailed (16).…”

Section: Introductionmentioning

confidence: 99%

The influence of the nature of the asparagine 289-linked oligosaccharide on the activation by urokinase and lysine binding properties of natural and recombinant human plasminogens.

Davidson¹

1993

J. Clin. Invest.

View full text Add to dashboard Cite

“…Heat treated virosomes were prepared by incubating the virosomes at 56°C for 20 min [10]. Degalactosylated virosomes were prepared by treating F-virosomes with clam exoglycosidase following the protocol described by Ghosh et al [11]. Blank virosomes were prepared by reconstituting Triton X-100 solubilised fraction in the absence of hygromycin B.…”

Section: Construction Of Loaded Virosomes and Fusogenic Liposomesmentioning

confidence: 99%

Targeted delivery of hygromycin B using reconstituted Sendai viral envelopes lacking hemagglutinin‐neuraminidase

Bagai

Sarkar

1993

FEBS Letters

View full text Add to dashboard Cite

Hygromycin B was encapsulated in reconstituted Sendai viral envelopes containing only the fusion (F) protein (F-virosomes). Incubation of loaded F-virosomes with cultured HepG2 cells resulted in fusion mediated delivery of hygromycin B to the cell cytoplasm, as was inferred from inhibition of DNA synthesis. Binding of the F-virosomes to HepG2 cells was mediated by the interaction of terminal fl-galactose residues of fusion protein with asialoglycoprotein receptor on HepG2 cells, subsequently leading to fusion between the two membranes. The cytotoxic effect of hygromycin B enclosed in F-virosomes was comparable with that of F, HN-virosomes containing both hemagglutinin-neuraminidase (HN) and F protein and F, HNred-virosomes containing HN whose disulfide bonds were irreversibly reduced (HNrJ. Hygromycin B loaded fusogenic liposomes were prepared by coreconstituting the viral envelope containing only fusion protein with exogenous lipids. These fusogenic liposomes were found to be more active than F-virosomes at the same fusion protein concentrations.

show abstract

Use of exoglycosidases from Mercenaria mercenaria (hard shelled clam) as a tool for structural studies of glycosphingolipids and glycoproteins

Cited by 11 publications

References 49 publications

Modification of the erythrocyte surface in rats bearing Yoshida ascites sarcoma is brought about by a tumour variant of α2-macroglobulin

Modification of the erythrocyte surface in rats bearing Yoshida ascites sarcoma is brought about by a tumour variant of α2-macroglobulin

The influence of the nature of the asparagine 289-linked oligosaccharide on the activation by urokinase and lysine binding properties of natural and recombinant human plasminogens.

Targeted delivery of hygromycin B using reconstituted Sendai viral envelopes lacking hemagglutinin‐neuraminidase

Contact Info

Product

Resources

About