Purification, properties, and partial structure elucidation of a high-molecular-weight glycoprotein from cervical mucus of the bonnet monkey (Macaca radiata)

Victor Bernard Hatcher, Günter O H Schwarzmann, Roger W. Jeanloz, Janet W. McArthur

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

A high-molecular-weight glycoprotein has been purified from the cervical mucus of the bonnet monkey (Macaca radiata). The glycoprotein was shown to be homogeneous by electrophoresis, sedimentation equilibrium, and N-terminal group determination, and to contain 19% protein, 19% D-galactose, 18% N-acetyl-D-galactosamine, 15% N-acetyl-D-glucosamine, 11% L-fucose, 10% sialic acid, and 1% sulfate groups, corresponding to about 1800 amino acid residues and 400 carbohydrate side chains of about 9 monosaccharides. The carbohydrate chains are linked to the peptide backbone through N-acetyl-D-galactosamine and serine (or threonine) residues. Reduction with dithiothreitol and alkylation with iodoacetic acid reduced the molecular mass from 1 to 0.5 × 106 daltons and produced subunits having the same size, charge. and N-terminal amino acid. Electrophoretic studies suggested the presence of disulfide bonds between two chains of the glycoprotein. Degradation with alkaline borohydride gave, after fractionation on Bio-Gel P-2, fractions containing L-fucose, D-galactose, N-acetyl-D-galactosaminitol, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, and sialic acid in the ratio of 1.0:3.0:1.0:1.0:1.3:1.0. Further fractionation by electrophoresis and paper chromatography gave a charged fraction representing 13% of the original glycoprotein. Enzymic degradation and methylation studies indicated the presence of the structure α-Gal-(1→3)-[Fuc(1-2)]-Gal-(1→4)-GlcNAc, linked to a core component containing N-acetyl-D-galactosaminitol.

Original languageEnglish (US)
Pages (from-to)1518-1524
Number of pages7
JournalBiochemistry
Volume16
Issue number7
StatePublished - 1977

Fingerprint

Macaca radiata
Cervix Mucus
Acetylgalactosamine
Purification
Glycoproteins
Molecular Weight
Molecular weight
Fucose
Acetylglucosamine
N-Acetylneuraminic Acid
Fractionation
Electrophoresis
Galactose
Carbohydrates
Iodoacetic Acid
Amino Acids
Degradation
Borohydrides
Paper Chromatography
Methylation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification, properties, and partial structure elucidation of a high-molecular-weight glycoprotein from cervical mucus of the bonnet monkey (Macaca radiata). / Hatcher, Victor Bernard; Schwarzmann, Günter O H; Jeanloz, Roger W.; McArthur, Janet W.

In: Biochemistry, Vol. 16, No. 7, 1977, p. 1518-1524.

Research output: Contribution to journalArticle

@article{f6e94446f6f74cbb8ad699b43d794a5f,
title = "Purification, properties, and partial structure elucidation of a high-molecular-weight glycoprotein from cervical mucus of the bonnet monkey (Macaca radiata)",
abstract = "A high-molecular-weight glycoprotein has been purified from the cervical mucus of the bonnet monkey (Macaca radiata). The glycoprotein was shown to be homogeneous by electrophoresis, sedimentation equilibrium, and N-terminal group determination, and to contain 19{\%} protein, 19{\%} D-galactose, 18{\%} N-acetyl-D-galactosamine, 15{\%} N-acetyl-D-glucosamine, 11{\%} L-fucose, 10{\%} sialic acid, and 1{\%} sulfate groups, corresponding to about 1800 amino acid residues and 400 carbohydrate side chains of about 9 monosaccharides. The carbohydrate chains are linked to the peptide backbone through N-acetyl-D-galactosamine and serine (or threonine) residues. Reduction with dithiothreitol and alkylation with iodoacetic acid reduced the molecular mass from 1 to 0.5 × 106 daltons and produced subunits having the same size, charge. and N-terminal amino acid. Electrophoretic studies suggested the presence of disulfide bonds between two chains of the glycoprotein. Degradation with alkaline borohydride gave, after fractionation on Bio-Gel P-2, fractions containing L-fucose, D-galactose, N-acetyl-D-galactosaminitol, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, and sialic acid in the ratio of 1.0:3.0:1.0:1.0:1.3:1.0. Further fractionation by electrophoresis and paper chromatography gave a charged fraction representing 13{\%} of the original glycoprotein. Enzymic degradation and methylation studies indicated the presence of the structure α-Gal-(1→3)-[Fuc(1-2)]-Gal-(1→4)-GlcNAc, linked to a core component containing N-acetyl-D-galactosaminitol.",
author = "Hatcher, {Victor Bernard} and Schwarzmann, {G{\"u}nter O H} and Jeanloz, {Roger W.} and McArthur, {Janet W.}",
year = "1977",
language = "English (US)",
volume = "16",
pages = "1518--1524",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "7",

}

TY - JOUR

T1 - Purification, properties, and partial structure elucidation of a high-molecular-weight glycoprotein from cervical mucus of the bonnet monkey (Macaca radiata)

AU - Hatcher, Victor Bernard

AU - Schwarzmann, Günter O H

AU - Jeanloz, Roger W.

AU - McArthur, Janet W.

PY - 1977

Y1 - 1977

N2 - A high-molecular-weight glycoprotein has been purified from the cervical mucus of the bonnet monkey (Macaca radiata). The glycoprotein was shown to be homogeneous by electrophoresis, sedimentation equilibrium, and N-terminal group determination, and to contain 19% protein, 19% D-galactose, 18% N-acetyl-D-galactosamine, 15% N-acetyl-D-glucosamine, 11% L-fucose, 10% sialic acid, and 1% sulfate groups, corresponding to about 1800 amino acid residues and 400 carbohydrate side chains of about 9 monosaccharides. The carbohydrate chains are linked to the peptide backbone through N-acetyl-D-galactosamine and serine (or threonine) residues. Reduction with dithiothreitol and alkylation with iodoacetic acid reduced the molecular mass from 1 to 0.5 × 106 daltons and produced subunits having the same size, charge. and N-terminal amino acid. Electrophoretic studies suggested the presence of disulfide bonds between two chains of the glycoprotein. Degradation with alkaline borohydride gave, after fractionation on Bio-Gel P-2, fractions containing L-fucose, D-galactose, N-acetyl-D-galactosaminitol, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, and sialic acid in the ratio of 1.0:3.0:1.0:1.0:1.3:1.0. Further fractionation by electrophoresis and paper chromatography gave a charged fraction representing 13% of the original glycoprotein. Enzymic degradation and methylation studies indicated the presence of the structure α-Gal-(1→3)-[Fuc(1-2)]-Gal-(1→4)-GlcNAc, linked to a core component containing N-acetyl-D-galactosaminitol.

AB - A high-molecular-weight glycoprotein has been purified from the cervical mucus of the bonnet monkey (Macaca radiata). The glycoprotein was shown to be homogeneous by electrophoresis, sedimentation equilibrium, and N-terminal group determination, and to contain 19% protein, 19% D-galactose, 18% N-acetyl-D-galactosamine, 15% N-acetyl-D-glucosamine, 11% L-fucose, 10% sialic acid, and 1% sulfate groups, corresponding to about 1800 amino acid residues and 400 carbohydrate side chains of about 9 monosaccharides. The carbohydrate chains are linked to the peptide backbone through N-acetyl-D-galactosamine and serine (or threonine) residues. Reduction with dithiothreitol and alkylation with iodoacetic acid reduced the molecular mass from 1 to 0.5 × 106 daltons and produced subunits having the same size, charge. and N-terminal amino acid. Electrophoretic studies suggested the presence of disulfide bonds between two chains of the glycoprotein. Degradation with alkaline borohydride gave, after fractionation on Bio-Gel P-2, fractions containing L-fucose, D-galactose, N-acetyl-D-galactosaminitol, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, and sialic acid in the ratio of 1.0:3.0:1.0:1.0:1.3:1.0. Further fractionation by electrophoresis and paper chromatography gave a charged fraction representing 13% of the original glycoprotein. Enzymic degradation and methylation studies indicated the presence of the structure α-Gal-(1→3)-[Fuc(1-2)]-Gal-(1→4)-GlcNAc, linked to a core component containing N-acetyl-D-galactosaminitol.

UR - http://www.scopus.com/inward/record.url?scp=0017344513&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017344513&partnerID=8YFLogxK

M3 - Article

VL - 16

SP - 1518

EP - 1524

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 7

ER -