Structures of O-linked oligosaccharides present in the proteoglycans secreted by human mammary epithelial cells

Channe Gowda, V. P. Bhavanandan, E. A. Davidson

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The structures of O-glycosidically linked oligosaccharides present in the heparan sulfate and chondroitin sulfate proteoglycans isolated from the culture medium of a normal (HBL-100) and a malignant (MDA-MB-231) human mammary epithelial cell line have been determined. Both proteoglycan types from the two cell lines contain a series of O-linked oligosaccharides ranging in size from di- to hexasaccharide. Cells were grown in the presence of either [3H]glucosamine or [3H]galactose and Na235SO4, and the proteoglycans were isolated as described (Gowda, D.C., Bhavanandan, V.P., and Davidson, E.A. (1986) J. Biol. Chem. 261, 4926-4934). The O-linked oligosaccharides were released from the proteoglycans by alkaline borohydride treatment and purified by a combination of gel filtration and high voltage paper electrophoresis. The structures of two neutral and seven acidic oligosaccharides were established based on sugar composition, the results of periodate oxidation, sequential exoglycosidase treatment, and methylation analysis. Periodate oxidation, taking advantage of tritium label at specific positions of constitutent sugars, proved to be a valuable tool in establishing the structure of isomeric components in the mixture. The oligosaccharide containing both sialic acid and ester sulfate is novel and has not been reported previously.

Original languageEnglish (US)
Pages (from-to)4935-4939
Number of pages5
JournalJournal of Biological Chemistry
Volume261
Issue number11
StatePublished - Dec 1 1986

Fingerprint

Proteoglycans
Oligosaccharides
Breast
Epithelial Cells
Sugars
Paper Electrophoresis
Chondroitin Sulfate Proteoglycans
Cell Line
Borohydrides
Oxidation
Methylation
Heparitin Sulfate
Tritium
Glycoside Hydrolases
Glucosamine
N-Acetylneuraminic Acid
Electrophoresis
Galactose
Sulfates
Gel Chromatography

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

@article{9f05c9db0e6a4588b2895c7c77cbbd3d,
title = "Structures of O-linked oligosaccharides present in the proteoglycans secreted by human mammary epithelial cells",
abstract = "The structures of O-glycosidically linked oligosaccharides present in the heparan sulfate and chondroitin sulfate proteoglycans isolated from the culture medium of a normal (HBL-100) and a malignant (MDA-MB-231) human mammary epithelial cell line have been determined. Both proteoglycan types from the two cell lines contain a series of O-linked oligosaccharides ranging in size from di- to hexasaccharide. Cells were grown in the presence of either [3H]glucosamine or [3H]galactose and Na235SO4, and the proteoglycans were isolated as described (Gowda, D.C., Bhavanandan, V.P., and Davidson, E.A. (1986) J. Biol. Chem. 261, 4926-4934). The O-linked oligosaccharides were released from the proteoglycans by alkaline borohydride treatment and purified by a combination of gel filtration and high voltage paper electrophoresis. The structures of two neutral and seven acidic oligosaccharides were established based on sugar composition, the results of periodate oxidation, sequential exoglycosidase treatment, and methylation analysis. Periodate oxidation, taking advantage of tritium label at specific positions of constitutent sugars, proved to be a valuable tool in establishing the structure of isomeric components in the mixture. The oligosaccharide containing both sialic acid and ester sulfate is novel and has not been reported previously.",
author = "Channe Gowda and Bhavanandan, {V. P.} and Davidson, {E. A.}",
year = "1986",
month = "12",
day = "1",
language = "English (US)",
volume = "261",
pages = "4935--4939",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "11",

}

Structures of O-linked oligosaccharides present in the proteoglycans secreted by human mammary epithelial cells. / Gowda, Channe; Bhavanandan, V. P.; Davidson, E. A.

In: Journal of Biological Chemistry, Vol. 261, No. 11, 01.12.1986, p. 4935-4939.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Structures of O-linked oligosaccharides present in the proteoglycans secreted by human mammary epithelial cells

AU - Gowda, Channe

AU - Bhavanandan, V. P.

AU - Davidson, E. A.

PY - 1986/12/1

Y1 - 1986/12/1

N2 - The structures of O-glycosidically linked oligosaccharides present in the heparan sulfate and chondroitin sulfate proteoglycans isolated from the culture medium of a normal (HBL-100) and a malignant (MDA-MB-231) human mammary epithelial cell line have been determined. Both proteoglycan types from the two cell lines contain a series of O-linked oligosaccharides ranging in size from di- to hexasaccharide. Cells were grown in the presence of either [3H]glucosamine or [3H]galactose and Na235SO4, and the proteoglycans were isolated as described (Gowda, D.C., Bhavanandan, V.P., and Davidson, E.A. (1986) J. Biol. Chem. 261, 4926-4934). The O-linked oligosaccharides were released from the proteoglycans by alkaline borohydride treatment and purified by a combination of gel filtration and high voltage paper electrophoresis. The structures of two neutral and seven acidic oligosaccharides were established based on sugar composition, the results of periodate oxidation, sequential exoglycosidase treatment, and methylation analysis. Periodate oxidation, taking advantage of tritium label at specific positions of constitutent sugars, proved to be a valuable tool in establishing the structure of isomeric components in the mixture. The oligosaccharide containing both sialic acid and ester sulfate is novel and has not been reported previously.

AB - The structures of O-glycosidically linked oligosaccharides present in the heparan sulfate and chondroitin sulfate proteoglycans isolated from the culture medium of a normal (HBL-100) and a malignant (MDA-MB-231) human mammary epithelial cell line have been determined. Both proteoglycan types from the two cell lines contain a series of O-linked oligosaccharides ranging in size from di- to hexasaccharide. Cells were grown in the presence of either [3H]glucosamine or [3H]galactose and Na235SO4, and the proteoglycans were isolated as described (Gowda, D.C., Bhavanandan, V.P., and Davidson, E.A. (1986) J. Biol. Chem. 261, 4926-4934). The O-linked oligosaccharides were released from the proteoglycans by alkaline borohydride treatment and purified by a combination of gel filtration and high voltage paper electrophoresis. The structures of two neutral and seven acidic oligosaccharides were established based on sugar composition, the results of periodate oxidation, sequential exoglycosidase treatment, and methylation analysis. Periodate oxidation, taking advantage of tritium label at specific positions of constitutent sugars, proved to be a valuable tool in establishing the structure of isomeric components in the mixture. The oligosaccharide containing both sialic acid and ester sulfate is novel and has not been reported previously.

UR - http://www.scopus.com/inward/record.url?scp=0022994160&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022994160&partnerID=8YFLogxK

M3 - Article

C2 - 3007481

AN - SCOPUS:0022994160

VL - 261

SP - 4935

EP - 4939

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 11

ER -