Biosynthesis of glycosphingolipids by human myeloid leukemia cells

Joanne Buehler, Eileen Qwan, Michael W. DeGregorio, Bruce A. Macher

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

We have performed comparative studies of the neutral glycosphingolipids synthesized by three human myeloid leukemia cell lines, K562, KG1, and HL-60, which were metabolically labeled with [14C]galactose, to evaluate changes in neutral glycosphingolipid synthesis with myeloid cell differentiation. Individual neutral glycosphingolipids containing one to four sugars were purified by a combination of the following methods: diethylaminoethyl-Sephadex column chromatography, acetylation-Florisil column chromatography, and high-performance liquid chromatography using an Iatrobead column. Compounds with one sugar were analyzed by thin-layer chromatography on borate plates. This analysis showed that HL-60 cells synthesize only glucosylceramide, whereas K562 and KG1 cells synthesize predominately glucosylceramide, but also a small amount of galactosylceramide. Compounds with two to four sugars were characterized by treatment with exo- and endoglycosidases. The results showed that K562 and KG1 cells are similar to cells from patients with acute leukemia in expressing two series (globo and neolacto) of natural glycosphingolipids, whereas the HL-60 cells are similar to mature human myeloid cells in expressing only one series (neolacto). Therefore, human myeloid leukemia cells blocked at different stages of differentiation vary in their ability to synthesize neutral glycosphingolipids.

Original languageEnglish (US)
Pages (from-to)6978-6984
Number of pages7
JournalBiochemistry
Volume24
Issue number24
StatePublished - 1985
Externally publishedYes

Fingerprint

Neutral Glycosphingolipids
Glycosphingolipids
Myeloid Leukemia
Biosynthesis
Myeloid Cells
Sugars
Glucosylceramides
Column chromatography
K562 Cells
Glycoside Hydrolases
HL-60 Cells
Chromatography
Galactosylceramides
Acetylation
Thin layer chromatography
Borates
High performance liquid chromatography
Thin Layer Chromatography
Galactose
Cell Differentiation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Buehler, J., Qwan, E., DeGregorio, M. W., & Macher, B. A. (1985). Biosynthesis of glycosphingolipids by human myeloid leukemia cells. Biochemistry, 24(24), 6978-6984.

Biosynthesis of glycosphingolipids by human myeloid leukemia cells. / Buehler, Joanne; Qwan, Eileen; DeGregorio, Michael W.; Macher, Bruce A.

In: Biochemistry, Vol. 24, No. 24, 1985, p. 6978-6984.

Research output: Contribution to journalArticle

Buehler, J, Qwan, E, DeGregorio, MW & Macher, BA 1985, 'Biosynthesis of glycosphingolipids by human myeloid leukemia cells', Biochemistry, vol. 24, no. 24, pp. 6978-6984.
Buehler J, Qwan E, DeGregorio MW, Macher BA. Biosynthesis of glycosphingolipids by human myeloid leukemia cells. Biochemistry. 1985;24(24):6978-6984.
Buehler, Joanne ; Qwan, Eileen ; DeGregorio, Michael W. ; Macher, Bruce A. / Biosynthesis of glycosphingolipids by human myeloid leukemia cells. In: Biochemistry. 1985 ; Vol. 24, No. 24. pp. 6978-6984.
@article{71ddb3d1a8ac45be93e0783fd827461f,
title = "Biosynthesis of glycosphingolipids by human myeloid leukemia cells",
abstract = "We have performed comparative studies of the neutral glycosphingolipids synthesized by three human myeloid leukemia cell lines, K562, KG1, and HL-60, which were metabolically labeled with [14C]galactose, to evaluate changes in neutral glycosphingolipid synthesis with myeloid cell differentiation. Individual neutral glycosphingolipids containing one to four sugars were purified by a combination of the following methods: diethylaminoethyl-Sephadex column chromatography, acetylation-Florisil column chromatography, and high-performance liquid chromatography using an Iatrobead column. Compounds with one sugar were analyzed by thin-layer chromatography on borate plates. This analysis showed that HL-60 cells synthesize only glucosylceramide, whereas K562 and KG1 cells synthesize predominately glucosylceramide, but also a small amount of galactosylceramide. Compounds with two to four sugars were characterized by treatment with exo- and endoglycosidases. The results showed that K562 and KG1 cells are similar to cells from patients with acute leukemia in expressing two series (globo and neolacto) of natural glycosphingolipids, whereas the HL-60 cells are similar to mature human myeloid cells in expressing only one series (neolacto). Therefore, human myeloid leukemia cells blocked at different stages of differentiation vary in their ability to synthesize neutral glycosphingolipids.",
author = "Joanne Buehler and Eileen Qwan and DeGregorio, {Michael W.} and Macher, {Bruce A.}",
year = "1985",
language = "English (US)",
volume = "24",
pages = "6978--6984",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "24",

}

TY - JOUR

T1 - Biosynthesis of glycosphingolipids by human myeloid leukemia cells

AU - Buehler, Joanne

AU - Qwan, Eileen

AU - DeGregorio, Michael W.

AU - Macher, Bruce A.

PY - 1985

Y1 - 1985

N2 - We have performed comparative studies of the neutral glycosphingolipids synthesized by three human myeloid leukemia cell lines, K562, KG1, and HL-60, which were metabolically labeled with [14C]galactose, to evaluate changes in neutral glycosphingolipid synthesis with myeloid cell differentiation. Individual neutral glycosphingolipids containing one to four sugars were purified by a combination of the following methods: diethylaminoethyl-Sephadex column chromatography, acetylation-Florisil column chromatography, and high-performance liquid chromatography using an Iatrobead column. Compounds with one sugar were analyzed by thin-layer chromatography on borate plates. This analysis showed that HL-60 cells synthesize only glucosylceramide, whereas K562 and KG1 cells synthesize predominately glucosylceramide, but also a small amount of galactosylceramide. Compounds with two to four sugars were characterized by treatment with exo- and endoglycosidases. The results showed that K562 and KG1 cells are similar to cells from patients with acute leukemia in expressing two series (globo and neolacto) of natural glycosphingolipids, whereas the HL-60 cells are similar to mature human myeloid cells in expressing only one series (neolacto). Therefore, human myeloid leukemia cells blocked at different stages of differentiation vary in their ability to synthesize neutral glycosphingolipids.

AB - We have performed comparative studies of the neutral glycosphingolipids synthesized by three human myeloid leukemia cell lines, K562, KG1, and HL-60, which were metabolically labeled with [14C]galactose, to evaluate changes in neutral glycosphingolipid synthesis with myeloid cell differentiation. Individual neutral glycosphingolipids containing one to four sugars were purified by a combination of the following methods: diethylaminoethyl-Sephadex column chromatography, acetylation-Florisil column chromatography, and high-performance liquid chromatography using an Iatrobead column. Compounds with one sugar were analyzed by thin-layer chromatography on borate plates. This analysis showed that HL-60 cells synthesize only glucosylceramide, whereas K562 and KG1 cells synthesize predominately glucosylceramide, but also a small amount of galactosylceramide. Compounds with two to four sugars were characterized by treatment with exo- and endoglycosidases. The results showed that K562 and KG1 cells are similar to cells from patients with acute leukemia in expressing two series (globo and neolacto) of natural glycosphingolipids, whereas the HL-60 cells are similar to mature human myeloid cells in expressing only one series (neolacto). Therefore, human myeloid leukemia cells blocked at different stages of differentiation vary in their ability to synthesize neutral glycosphingolipids.

UR - http://www.scopus.com/inward/record.url?scp=0022404458&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022404458&partnerID=8YFLogxK

M3 - Article

VL - 24

SP - 6978

EP - 6984

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 24

ER -