Core sugar residues of the N-linked oligosaccharides of Russell's viper venom factor X-activator maintain functionally active polypeptide structure

Channe Gowda, Craig M. Jackson, Gary P. Kurzban, Peter McPhie, Eugene A. Davidson

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

We previously showed that the factor X activator of Russell's viper venom (RVV-X) contains six N-linked oligosaccharide chains: four in the heavy chain and one in each of the two light chains [Gowda, D. C., Jackson, C. M., Hensley, P., and Davidson, E. A. (1994) J. Biol. Chem. 269, 1064410650).] In the present study, we have investigated the role of the carbohydrate moieties in the structure and functional activity of RVV-X. Sequential removal of sugar residues from the terminal ends by exoglycosidases, up to 50% of total carbohydrates, did not significantly alter the activity of RVV-X, demonstrating that the peripheral carbohydrate moieties are not involved in interactions with factor X. However, removal of whole oligosaccharide chains by N-glycanase caused an almost total loss of the ability of RVV-X to activate factor X to factor X(a). In parallel with these observations, circular dichroism spectroscopy showed that complete deglycosylation, but not the removal of peripheral sugars, caused a significant change in the secondary structure. Together, these data demonstrate that the oligosaccharide chains are necessary for the functional activity, and that the trimannosylchitobiose core residues are sufficient for the maintenance of the native polypeptide structure.

Original languageEnglish (US)
Pages (from-to)5833-5837
Number of pages5
JournalBiochemistry
Volume35
Issue number18
DOIs
StatePublished - May 7 1996

Fingerprint

Oligosaccharides
Sugars
Factor X
Peptides
Carbohydrates
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Circular dichroism spectroscopy
Glycoside Hydrolases
Circular Dichroism
Spectrum Analysis
Maintenance
russellysin
Light

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Gowda, Channe ; Jackson, Craig M. ; Kurzban, Gary P. ; McPhie, Peter ; Davidson, Eugene A. / Core sugar residues of the N-linked oligosaccharides of Russell's viper venom factor X-activator maintain functionally active polypeptide structure. In: Biochemistry. 1996 ; Vol. 35, No. 18. pp. 5833-5837.
@article{93ee6e93c18045a39d7af65a4678034c,
title = "Core sugar residues of the N-linked oligosaccharides of Russell's viper venom factor X-activator maintain functionally active polypeptide structure",
abstract = "We previously showed that the factor X activator of Russell's viper venom (RVV-X) contains six N-linked oligosaccharide chains: four in the heavy chain and one in each of the two light chains [Gowda, D. C., Jackson, C. M., Hensley, P., and Davidson, E. A. (1994) J. Biol. Chem. 269, 1064410650).] In the present study, we have investigated the role of the carbohydrate moieties in the structure and functional activity of RVV-X. Sequential removal of sugar residues from the terminal ends by exoglycosidases, up to 50{\%} of total carbohydrates, did not significantly alter the activity of RVV-X, demonstrating that the peripheral carbohydrate moieties are not involved in interactions with factor X. However, removal of whole oligosaccharide chains by N-glycanase caused an almost total loss of the ability of RVV-X to activate factor X to factor X(a). In parallel with these observations, circular dichroism spectroscopy showed that complete deglycosylation, but not the removal of peripheral sugars, caused a significant change in the secondary structure. Together, these data demonstrate that the oligosaccharide chains are necessary for the functional activity, and that the trimannosylchitobiose core residues are sufficient for the maintenance of the native polypeptide structure.",
author = "Channe Gowda and Jackson, {Craig M.} and Kurzban, {Gary P.} and Peter McPhie and Davidson, {Eugene A.}",
year = "1996",
month = "5",
day = "7",
doi = "10.1021/bi953043e",
language = "English (US)",
volume = "35",
pages = "5833--5837",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "18",

}

Core sugar residues of the N-linked oligosaccharides of Russell's viper venom factor X-activator maintain functionally active polypeptide structure. / Gowda, Channe; Jackson, Craig M.; Kurzban, Gary P.; McPhie, Peter; Davidson, Eugene A.

In: Biochemistry, Vol. 35, No. 18, 07.05.1996, p. 5833-5837.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Core sugar residues of the N-linked oligosaccharides of Russell's viper venom factor X-activator maintain functionally active polypeptide structure

AU - Gowda, Channe

AU - Jackson, Craig M.

AU - Kurzban, Gary P.

AU - McPhie, Peter

AU - Davidson, Eugene A.

PY - 1996/5/7

Y1 - 1996/5/7

N2 - We previously showed that the factor X activator of Russell's viper venom (RVV-X) contains six N-linked oligosaccharide chains: four in the heavy chain and one in each of the two light chains [Gowda, D. C., Jackson, C. M., Hensley, P., and Davidson, E. A. (1994) J. Biol. Chem. 269, 1064410650).] In the present study, we have investigated the role of the carbohydrate moieties in the structure and functional activity of RVV-X. Sequential removal of sugar residues from the terminal ends by exoglycosidases, up to 50% of total carbohydrates, did not significantly alter the activity of RVV-X, demonstrating that the peripheral carbohydrate moieties are not involved in interactions with factor X. However, removal of whole oligosaccharide chains by N-glycanase caused an almost total loss of the ability of RVV-X to activate factor X to factor X(a). In parallel with these observations, circular dichroism spectroscopy showed that complete deglycosylation, but not the removal of peripheral sugars, caused a significant change in the secondary structure. Together, these data demonstrate that the oligosaccharide chains are necessary for the functional activity, and that the trimannosylchitobiose core residues are sufficient for the maintenance of the native polypeptide structure.

AB - We previously showed that the factor X activator of Russell's viper venom (RVV-X) contains six N-linked oligosaccharide chains: four in the heavy chain and one in each of the two light chains [Gowda, D. C., Jackson, C. M., Hensley, P., and Davidson, E. A. (1994) J. Biol. Chem. 269, 1064410650).] In the present study, we have investigated the role of the carbohydrate moieties in the structure and functional activity of RVV-X. Sequential removal of sugar residues from the terminal ends by exoglycosidases, up to 50% of total carbohydrates, did not significantly alter the activity of RVV-X, demonstrating that the peripheral carbohydrate moieties are not involved in interactions with factor X. However, removal of whole oligosaccharide chains by N-glycanase caused an almost total loss of the ability of RVV-X to activate factor X to factor X(a). In parallel with these observations, circular dichroism spectroscopy showed that complete deglycosylation, but not the removal of peripheral sugars, caused a significant change in the secondary structure. Together, these data demonstrate that the oligosaccharide chains are necessary for the functional activity, and that the trimannosylchitobiose core residues are sufficient for the maintenance of the native polypeptide structure.

UR - http://www.scopus.com/inward/record.url?scp=0029971864&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029971864&partnerID=8YFLogxK

U2 - 10.1021/bi953043e

DO - 10.1021/bi953043e

M3 - Article

VL - 35

SP - 5833

EP - 5837

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 18

ER -