Lipid Composition of PC 12 Pheochromocytoma Cells: Characterization of Globoside as a Major Neutral Glycolipid

Toshio Ariga, Lawrence J. Macala, Megumi Saito, Renee K. Margolis, Lloyd A. Greene, Richard U. Margolis, Robert K Yu

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

We have studied the lipid composition of PC 12 pheochromocytoma cells cultured in the presence and absence of nerve growth factor (NGF). Neutral and acidic lipid fractions were isolated by column chromatography on DEAE-Sephadex and analyzed by high-performance thin-layer chromatography (HPTLC). The total lipid concentration was approximately 220 jug/mg of protein, and the concentration of neutral glycolipids was 1.6-1.8 Mg/mg of protein for both NGF-treated and untreated cells. The neutral glycolipid fraction contained a major component, which accounted for approximately 80% of the total and which was characterized as globoside on the basis of HPTLC mobility, carbohydrate analysis, fast atom bombardment mass spectrometry, and mild acid hydrolysis. The major fatty acids of globoside were Cl6:0 (10%), C18:0 (16%), C22:0 (23%), C24:l (17%), and C24:0 (24%). C18 sphingenine accounted for almost all of the long-chain bases. The other neutral glycolipids were tentatively identified as glucosylceramide (15%), lactosylceramide (4%), and globotriosylceramide (4.5%). The concentration of ganglioside sialic acid was approximately 0.34 and 0.18 p%f mg of protein for cells grown in the presence and absence of NGF, respectively. Although there was an increase in ganglioside concentration in NGF-treated cells, NGF did not produce any differential effects on the relative proportions of the individual gangliosides. Several of the gangliosides appear to contain fucose, and one of these was tentatively identified as fucosyl-GMltBrain-type gangliosides of the ganglio series were also detected by an HPTLC-immunostaining method. However, the fatty acid and long chain base compositions of PC 12 cell gangliosides (and their TLC mobility) differ from those of brain gangliosides. The major lipids of PC 12 cells, which were not significantly affected by NGF treatment, were phosphatidylcholine (40%), phosphatidylethanolamine (22%), and cholesterol (13%).

Original languageEnglish (US)
Pages (from-to)52-58
Number of pages7
JournalBiochemistry
Volume27
Issue number1
DOIs
StatePublished - Jan 1 1988
Externally publishedYes

Fingerprint

Globosides
Gangliosides
Glycolipids
Pheochromocytoma
Nerve Growth Factor
Lipids
Thin layer chromatography
Chemical analysis
Thin Layer Chromatography
Fatty Acids
Cells
Glucosylceramides
Fast Atom Bombardment Mass Spectrometry
DEAE-Dextran
Column chromatography
Proteins
Fucose
Base Composition
N-Acetylneuraminic Acid
Phosphatidylcholines

ASJC Scopus subject areas

  • Biochemistry

Cite this

Lipid Composition of PC 12 Pheochromocytoma Cells : Characterization of Globoside as a Major Neutral Glycolipid. / Ariga, Toshio; Macala, Lawrence J.; Saito, Megumi; Margolis, Renee K.; Greene, Lloyd A.; Margolis, Richard U.; Yu, Robert K.

In: Biochemistry, Vol. 27, No. 1, 01.01.1988, p. 52-58.

Research output: Contribution to journalArticle

Ariga, Toshio ; Macala, Lawrence J. ; Saito, Megumi ; Margolis, Renee K. ; Greene, Lloyd A. ; Margolis, Richard U. ; Yu, Robert K. / Lipid Composition of PC 12 Pheochromocytoma Cells : Characterization of Globoside as a Major Neutral Glycolipid. In: Biochemistry. 1988 ; Vol. 27, No. 1. pp. 52-58.
@article{da68ff1dd6b94f1781dbaf1b937efcf2,
title = "Lipid Composition of PC 12 Pheochromocytoma Cells: Characterization of Globoside as a Major Neutral Glycolipid",
abstract = "We have studied the lipid composition of PC 12 pheochromocytoma cells cultured in the presence and absence of nerve growth factor (NGF). Neutral and acidic lipid fractions were isolated by column chromatography on DEAE-Sephadex and analyzed by high-performance thin-layer chromatography (HPTLC). The total lipid concentration was approximately 220 jug/mg of protein, and the concentration of neutral glycolipids was 1.6-1.8 Mg/mg of protein for both NGF-treated and untreated cells. The neutral glycolipid fraction contained a major component, which accounted for approximately 80{\%} of the total and which was characterized as globoside on the basis of HPTLC mobility, carbohydrate analysis, fast atom bombardment mass spectrometry, and mild acid hydrolysis. The major fatty acids of globoside were Cl6:0 (10{\%}), C18:0 (16{\%}), C22:0 (23{\%}), C24:l (17{\%}), and C24:0 (24{\%}). C18 sphingenine accounted for almost all of the long-chain bases. The other neutral glycolipids were tentatively identified as glucosylceramide (15{\%}), lactosylceramide (4{\%}), and globotriosylceramide (4.5{\%}). The concentration of ganglioside sialic acid was approximately 0.34 and 0.18 p{\%}f mg of protein for cells grown in the presence and absence of NGF, respectively. Although there was an increase in ganglioside concentration in NGF-treated cells, NGF did not produce any differential effects on the relative proportions of the individual gangliosides. Several of the gangliosides appear to contain fucose, and one of these was tentatively identified as fucosyl-GMltBrain-type gangliosides of the ganglio series were also detected by an HPTLC-immunostaining method. However, the fatty acid and long chain base compositions of PC 12 cell gangliosides (and their TLC mobility) differ from those of brain gangliosides. The major lipids of PC 12 cells, which were not significantly affected by NGF treatment, were phosphatidylcholine (40{\%}), phosphatidylethanolamine (22{\%}), and cholesterol (13{\%}).",
author = "Toshio Ariga and Macala, {Lawrence J.} and Megumi Saito and Margolis, {Renee K.} and Greene, {Lloyd A.} and Margolis, {Richard U.} and Yu, {Robert K}",
year = "1988",
month = "1",
day = "1",
doi = "10.1021/bi00401a010",
language = "English (US)",
volume = "27",
pages = "52--58",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "1",

}

TY - JOUR

T1 - Lipid Composition of PC 12 Pheochromocytoma Cells

T2 - Characterization of Globoside as a Major Neutral Glycolipid

AU - Ariga, Toshio

AU - Macala, Lawrence J.

AU - Saito, Megumi

AU - Margolis, Renee K.

AU - Greene, Lloyd A.

AU - Margolis, Richard U.

AU - Yu, Robert K

PY - 1988/1/1

Y1 - 1988/1/1

N2 - We have studied the lipid composition of PC 12 pheochromocytoma cells cultured in the presence and absence of nerve growth factor (NGF). Neutral and acidic lipid fractions were isolated by column chromatography on DEAE-Sephadex and analyzed by high-performance thin-layer chromatography (HPTLC). The total lipid concentration was approximately 220 jug/mg of protein, and the concentration of neutral glycolipids was 1.6-1.8 Mg/mg of protein for both NGF-treated and untreated cells. The neutral glycolipid fraction contained a major component, which accounted for approximately 80% of the total and which was characterized as globoside on the basis of HPTLC mobility, carbohydrate analysis, fast atom bombardment mass spectrometry, and mild acid hydrolysis. The major fatty acids of globoside were Cl6:0 (10%), C18:0 (16%), C22:0 (23%), C24:l (17%), and C24:0 (24%). C18 sphingenine accounted for almost all of the long-chain bases. The other neutral glycolipids were tentatively identified as glucosylceramide (15%), lactosylceramide (4%), and globotriosylceramide (4.5%). The concentration of ganglioside sialic acid was approximately 0.34 and 0.18 p%f mg of protein for cells grown in the presence and absence of NGF, respectively. Although there was an increase in ganglioside concentration in NGF-treated cells, NGF did not produce any differential effects on the relative proportions of the individual gangliosides. Several of the gangliosides appear to contain fucose, and one of these was tentatively identified as fucosyl-GMltBrain-type gangliosides of the ganglio series were also detected by an HPTLC-immunostaining method. However, the fatty acid and long chain base compositions of PC 12 cell gangliosides (and their TLC mobility) differ from those of brain gangliosides. The major lipids of PC 12 cells, which were not significantly affected by NGF treatment, were phosphatidylcholine (40%), phosphatidylethanolamine (22%), and cholesterol (13%).

AB - We have studied the lipid composition of PC 12 pheochromocytoma cells cultured in the presence and absence of nerve growth factor (NGF). Neutral and acidic lipid fractions were isolated by column chromatography on DEAE-Sephadex and analyzed by high-performance thin-layer chromatography (HPTLC). The total lipid concentration was approximately 220 jug/mg of protein, and the concentration of neutral glycolipids was 1.6-1.8 Mg/mg of protein for both NGF-treated and untreated cells. The neutral glycolipid fraction contained a major component, which accounted for approximately 80% of the total and which was characterized as globoside on the basis of HPTLC mobility, carbohydrate analysis, fast atom bombardment mass spectrometry, and mild acid hydrolysis. The major fatty acids of globoside were Cl6:0 (10%), C18:0 (16%), C22:0 (23%), C24:l (17%), and C24:0 (24%). C18 sphingenine accounted for almost all of the long-chain bases. The other neutral glycolipids were tentatively identified as glucosylceramide (15%), lactosylceramide (4%), and globotriosylceramide (4.5%). The concentration of ganglioside sialic acid was approximately 0.34 and 0.18 p%f mg of protein for cells grown in the presence and absence of NGF, respectively. Although there was an increase in ganglioside concentration in NGF-treated cells, NGF did not produce any differential effects on the relative proportions of the individual gangliosides. Several of the gangliosides appear to contain fucose, and one of these was tentatively identified as fucosyl-GMltBrain-type gangliosides of the ganglio series were also detected by an HPTLC-immunostaining method. However, the fatty acid and long chain base compositions of PC 12 cell gangliosides (and their TLC mobility) differ from those of brain gangliosides. The major lipids of PC 12 cells, which were not significantly affected by NGF treatment, were phosphatidylcholine (40%), phosphatidylethanolamine (22%), and cholesterol (13%).

UR - http://www.scopus.com/inward/record.url?scp=0023872609&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023872609&partnerID=8YFLogxK

U2 - 10.1021/bi00401a010

DO - 10.1021/bi00401a010

M3 - Article

C2 - 3349045

AN - SCOPUS:0023872609

VL - 27

SP - 52

EP - 58

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 1

ER -