Mucin biosynthesis. Properties of a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase

P. W. Cheng, W. E. Wingert, M. R. Little, R. Wei

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

We have characterized a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase that catalyses the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to the C-6 of the N-acetylgalactosamine residue of galactosyl-β1→3-N-acetylgalactosamine. Optimal enzyme activity was obtained between pH 7.5-8.5, at 5 mM-MnCl2, and at 0.06-0.08% (v/v) Triton X-100 (or Nonidet P-40), or 0.5-5.0% (v/v) Tween 20. Ba2+, Mg2+ and Ca2+ could partially replace Mn2+, but Co2+, Fe2+, Cd2+ and Zn2+ could not. Sodium dodecyl sulphate, cetylpyridinium chloride, sodium deoxycholate, octyl β-D-glucoside, digitonin and alkyl alcohols were less effective in enhancing enzyme activity, and dimethyl sulphoxide was ineffective. The apparent Michaelis constants were 1.25 mM for UDP-N-acetylglucosamine, 0.94-3.34 mM for freezing-point-depressing glycoprotein and 0.19 mM for periodate-treated blood-group-A porcine submaxillary mucin. Asialo ovine submaxillary mucin could not serve as the glycosyl acceptor. The structure of the 14C-labelled oligosaccharide obtained by alkaline-borohydride treatment of the product was identified as Galβ1→3(Glc-NAcβ1→6)N-acetylgalactosaminitol by β-hexosaminidase treatment, gas chromatography-mass spectrometry and 1H-n.m.r. (270 MHz) analysis. The enzyme is important in the regulation of mucin oligosaccharide biosynthesis.

Original languageEnglish (US)
Pages (from-to)405-412
Number of pages8
JournalBiochemical Journal
Volume227
Issue number2
DOIs
StatePublished - Jan 1 1985

Fingerprint

Mucin-6
Biosynthesis
Mucins
Uridine Diphosphate N-Acetylglucosamine
Acetylgalactosamine
Acetylglucosamine
Enzyme activity
Oligosaccharides
Enzymes
Cetylpyridinium
Hexosaminidases
Digitonin
Borohydrides
Deoxycholic Acid
Uridine Diphosphate
Polysorbates
Octoxynol
Glucosides
Blood Group Antigens
Dimethyl Sulfoxide

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Mucin biosynthesis. Properties of a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase. / Cheng, P. W.; Wingert, W. E.; Little, M. R.; Wei, R.

In: Biochemical Journal, Vol. 227, No. 2, 01.01.1985, p. 405-412.

Research output: Contribution to journalArticle

Cheng, P. W. ; Wingert, W. E. ; Little, M. R. ; Wei, R. / Mucin biosynthesis. Properties of a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase. In: Biochemical Journal. 1985 ; Vol. 227, No. 2. pp. 405-412.
@article{bef4d340eb2c4ac09d26e43474da3f4f,
title = "Mucin biosynthesis. Properties of a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase",
abstract = "We have characterized a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase that catalyses the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to the C-6 of the N-acetylgalactosamine residue of galactosyl-β1→3-N-acetylgalactosamine. Optimal enzyme activity was obtained between pH 7.5-8.5, at 5 mM-MnCl2, and at 0.06-0.08{\%} (v/v) Triton X-100 (or Nonidet P-40), or 0.5-5.0{\%} (v/v) Tween 20. Ba2+, Mg2+ and Ca2+ could partially replace Mn2+, but Co2+, Fe2+, Cd2+ and Zn2+ could not. Sodium dodecyl sulphate, cetylpyridinium chloride, sodium deoxycholate, octyl β-D-glucoside, digitonin and alkyl alcohols were less effective in enhancing enzyme activity, and dimethyl sulphoxide was ineffective. The apparent Michaelis constants were 1.25 mM for UDP-N-acetylglucosamine, 0.94-3.34 mM for freezing-point-depressing glycoprotein and 0.19 mM for periodate-treated blood-group-A porcine submaxillary mucin. Asialo ovine submaxillary mucin could not serve as the glycosyl acceptor. The structure of the 14C-labelled oligosaccharide obtained by alkaline-borohydride treatment of the product was identified as Galβ1→3(Glc-NAcβ1→6)N-acetylgalactosaminitol by β-hexosaminidase treatment, gas chromatography-mass spectrometry and 1H-n.m.r. (270 MHz) analysis. The enzyme is important in the regulation of mucin oligosaccharide biosynthesis.",
author = "Cheng, {P. W.} and Wingert, {W. E.} and Little, {M. R.} and R. Wei",
year = "1985",
month = "1",
day = "1",
doi = "10.1042/bj2270405",
language = "English (US)",
volume = "227",
pages = "405--412",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

TY - JOUR

T1 - Mucin biosynthesis. Properties of a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase

AU - Cheng, P. W.

AU - Wingert, W. E.

AU - Little, M. R.

AU - Wei, R.

PY - 1985/1/1

Y1 - 1985/1/1

N2 - We have characterized a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase that catalyses the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to the C-6 of the N-acetylgalactosamine residue of galactosyl-β1→3-N-acetylgalactosamine. Optimal enzyme activity was obtained between pH 7.5-8.5, at 5 mM-MnCl2, and at 0.06-0.08% (v/v) Triton X-100 (or Nonidet P-40), or 0.5-5.0% (v/v) Tween 20. Ba2+, Mg2+ and Ca2+ could partially replace Mn2+, but Co2+, Fe2+, Cd2+ and Zn2+ could not. Sodium dodecyl sulphate, cetylpyridinium chloride, sodium deoxycholate, octyl β-D-glucoside, digitonin and alkyl alcohols were less effective in enhancing enzyme activity, and dimethyl sulphoxide was ineffective. The apparent Michaelis constants were 1.25 mM for UDP-N-acetylglucosamine, 0.94-3.34 mM for freezing-point-depressing glycoprotein and 0.19 mM for periodate-treated blood-group-A porcine submaxillary mucin. Asialo ovine submaxillary mucin could not serve as the glycosyl acceptor. The structure of the 14C-labelled oligosaccharide obtained by alkaline-borohydride treatment of the product was identified as Galβ1→3(Glc-NAcβ1→6)N-acetylgalactosaminitol by β-hexosaminidase treatment, gas chromatography-mass spectrometry and 1H-n.m.r. (270 MHz) analysis. The enzyme is important in the regulation of mucin oligosaccharide biosynthesis.

AB - We have characterized a bovine tracheal mucin β-6-N-acetylglucosaminyltransferase that catalyses the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to the C-6 of the N-acetylgalactosamine residue of galactosyl-β1→3-N-acetylgalactosamine. Optimal enzyme activity was obtained between pH 7.5-8.5, at 5 mM-MnCl2, and at 0.06-0.08% (v/v) Triton X-100 (or Nonidet P-40), or 0.5-5.0% (v/v) Tween 20. Ba2+, Mg2+ and Ca2+ could partially replace Mn2+, but Co2+, Fe2+, Cd2+ and Zn2+ could not. Sodium dodecyl sulphate, cetylpyridinium chloride, sodium deoxycholate, octyl β-D-glucoside, digitonin and alkyl alcohols were less effective in enhancing enzyme activity, and dimethyl sulphoxide was ineffective. The apparent Michaelis constants were 1.25 mM for UDP-N-acetylglucosamine, 0.94-3.34 mM for freezing-point-depressing glycoprotein and 0.19 mM for periodate-treated blood-group-A porcine submaxillary mucin. Asialo ovine submaxillary mucin could not serve as the glycosyl acceptor. The structure of the 14C-labelled oligosaccharide obtained by alkaline-borohydride treatment of the product was identified as Galβ1→3(Glc-NAcβ1→6)N-acetylgalactosaminitol by β-hexosaminidase treatment, gas chromatography-mass spectrometry and 1H-n.m.r. (270 MHz) analysis. The enzyme is important in the regulation of mucin oligosaccharide biosynthesis.

UR - http://www.scopus.com/inward/record.url?scp=0021793957&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021793957&partnerID=8YFLogxK

U2 - 10.1042/bj2270405

DO - 10.1042/bj2270405

M3 - Article

C2 - 3924025

AN - SCOPUS:0021793957

VL - 227

SP - 405

EP - 412

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -