UDP-galactose

globotriaosylceramide α-galactosyltransferase activity in rat pheochromocytoma (PC12h) cells

S. Pal, M. Saito, T. Ariga, Robert K Yu

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The activity of α-galactosyltransferase in cultured rat pheochromocytoma subcloned (PC12h) cells was examined using Gb3 as the acceptor for the galactose from UDP-galactose. The major reaction product was identified as galα1-3Gb3 based on its mobility on thin-layer chromatographic (TLC) plates and susceptibility to specific galactosidases. The enzyme activity in PC12h cells was the highest at pH 7.0 while the presence of Triton CF-54 (0.1%) and Mn2+ (5 mM) was required for its full activity. The apparent K(m) values for Gb3 and UDP-galactose were 57 and 17 μM, respectively. The enzyme activity in PC12h cells was compared with that in parent PC12 cells, in which galα1-3Gb3 is not expressed in an appreciable amount. In the enzyme reaction with exogenous Gb3, the enzyme activity in PC12h cells was about 1.5-fold higher than that in PC12 cells. In the absence of exogenous Gb3, this difference became even more pronounced; galα1-3Gb3 was generated from endogenous Gb3 at a much higher rate in PC12h cells than in PC12 cells. These findings suggest that the higher level of the α-galactosyltransferase activity in PC12h cells may, at least in part, be responsible for the accumulation of unique neutral glycosphingolipids having galα1-3 terminal residues in the cells.

Original languageEnglish (US)
Pages (from-to)411-417
Number of pages7
JournalJournal of Lipid Research
Volume33
Issue number3
StatePublished - Jan 1 1992
Externally publishedYes

Fingerprint

Uridine Diphosphate Galactose
Galactosyltransferases
Enzyme activity
Pheochromocytoma
Rats
Neutral Glycosphingolipids
PC12 Cells
Galactosidases
Reaction products
Enzymes
globotriaosylceramide
Galactose

Keywords

  • differentiation
  • galα1-3Gb3
  • glycosphingolipids

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

UDP-galactose : globotriaosylceramide α-galactosyltransferase activity in rat pheochromocytoma (PC12h) cells. / Pal, S.; Saito, M.; Ariga, T.; Yu, Robert K.

In: Journal of Lipid Research, Vol. 33, No. 3, 01.01.1992, p. 411-417.

Research output: Contribution to journalArticle

@article{90e5df7347c24db19589cfa735c163bb,
title = "UDP-galactose: globotriaosylceramide α-galactosyltransferase activity in rat pheochromocytoma (PC12h) cells",
abstract = "The activity of α-galactosyltransferase in cultured rat pheochromocytoma subcloned (PC12h) cells was examined using Gb3 as the acceptor for the galactose from UDP-galactose. The major reaction product was identified as galα1-3Gb3 based on its mobility on thin-layer chromatographic (TLC) plates and susceptibility to specific galactosidases. The enzyme activity in PC12h cells was the highest at pH 7.0 while the presence of Triton CF-54 (0.1{\%}) and Mn2+ (5 mM) was required for its full activity. The apparent K(m) values for Gb3 and UDP-galactose were 57 and 17 μM, respectively. The enzyme activity in PC12h cells was compared with that in parent PC12 cells, in which galα1-3Gb3 is not expressed in an appreciable amount. In the enzyme reaction with exogenous Gb3, the enzyme activity in PC12h cells was about 1.5-fold higher than that in PC12 cells. In the absence of exogenous Gb3, this difference became even more pronounced; galα1-3Gb3 was generated from endogenous Gb3 at a much higher rate in PC12h cells than in PC12 cells. These findings suggest that the higher level of the α-galactosyltransferase activity in PC12h cells may, at least in part, be responsible for the accumulation of unique neutral glycosphingolipids having galα1-3 terminal residues in the cells.",
keywords = "differentiation, galα1-3Gb3, glycosphingolipids",
author = "S. Pal and M. Saito and T. Ariga and Yu, {Robert K}",
year = "1992",
month = "1",
day = "1",
language = "English (US)",
volume = "33",
pages = "411--417",
journal = "Journal of Lipid Research",
issn = "0022-2275",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "3",

}

TY - JOUR

T1 - UDP-galactose

T2 - globotriaosylceramide α-galactosyltransferase activity in rat pheochromocytoma (PC12h) cells

AU - Pal, S.

AU - Saito, M.

AU - Ariga, T.

AU - Yu, Robert K

PY - 1992/1/1

Y1 - 1992/1/1

N2 - The activity of α-galactosyltransferase in cultured rat pheochromocytoma subcloned (PC12h) cells was examined using Gb3 as the acceptor for the galactose from UDP-galactose. The major reaction product was identified as galα1-3Gb3 based on its mobility on thin-layer chromatographic (TLC) plates and susceptibility to specific galactosidases. The enzyme activity in PC12h cells was the highest at pH 7.0 while the presence of Triton CF-54 (0.1%) and Mn2+ (5 mM) was required for its full activity. The apparent K(m) values for Gb3 and UDP-galactose were 57 and 17 μM, respectively. The enzyme activity in PC12h cells was compared with that in parent PC12 cells, in which galα1-3Gb3 is not expressed in an appreciable amount. In the enzyme reaction with exogenous Gb3, the enzyme activity in PC12h cells was about 1.5-fold higher than that in PC12 cells. In the absence of exogenous Gb3, this difference became even more pronounced; galα1-3Gb3 was generated from endogenous Gb3 at a much higher rate in PC12h cells than in PC12 cells. These findings suggest that the higher level of the α-galactosyltransferase activity in PC12h cells may, at least in part, be responsible for the accumulation of unique neutral glycosphingolipids having galα1-3 terminal residues in the cells.

AB - The activity of α-galactosyltransferase in cultured rat pheochromocytoma subcloned (PC12h) cells was examined using Gb3 as the acceptor for the galactose from UDP-galactose. The major reaction product was identified as galα1-3Gb3 based on its mobility on thin-layer chromatographic (TLC) plates and susceptibility to specific galactosidases. The enzyme activity in PC12h cells was the highest at pH 7.0 while the presence of Triton CF-54 (0.1%) and Mn2+ (5 mM) was required for its full activity. The apparent K(m) values for Gb3 and UDP-galactose were 57 and 17 μM, respectively. The enzyme activity in PC12h cells was compared with that in parent PC12 cells, in which galα1-3Gb3 is not expressed in an appreciable amount. In the enzyme reaction with exogenous Gb3, the enzyme activity in PC12h cells was about 1.5-fold higher than that in PC12 cells. In the absence of exogenous Gb3, this difference became even more pronounced; galα1-3Gb3 was generated from endogenous Gb3 at a much higher rate in PC12h cells than in PC12 cells. These findings suggest that the higher level of the α-galactosyltransferase activity in PC12h cells may, at least in part, be responsible for the accumulation of unique neutral glycosphingolipids having galα1-3 terminal residues in the cells.

KW - differentiation

KW - galα1-3Gb3

KW - glycosphingolipids

UR - http://www.scopus.com/inward/record.url?scp=0026596121&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026596121&partnerID=8YFLogxK

M3 - Article

VL - 33

SP - 411

EP - 417

JO - Journal of Lipid Research

JF - Journal of Lipid Research

SN - 0022-2275

IS - 3

ER -