Detergent-free domain isolated from Xenopus egg plasma membrane with properties similar to those of detergent-resistant membranes

Ayala Luria, Vaida Vegelyte-Avery, Brad Stith, Nelly M. Tsvetkova, Willem F. Wolkers, John H. Crowe, Fern Tablin, Richard Nuccitelli

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Microdomains known as "rafts" have been isolated from many cell types as detergent-resistant membranes (DRMs) and are enriched in sphingolipids and cholesterol. However, there has been considerable controversy over whether such domains are found in native membranes or are artificially generated by the purification procedure. This controversy is based at least in part on the fact that raft membranes were first detected following detergent extraction in the cold. We isolated two plasma membrane fractions, without detergent treatment, using a discontinuous sucrose density gradient. One fraction was designated "light" and the other "heavy." These fractions were compared with DRMs, which were isolated in the presence of 1% Triton X-100. We found that Xenopus DRMs are enriched with sphingomyelin and cholesterol and exhibit a phase state similar to the liquid-ordered phase. Comparison of DRM complexes with the light and heavy plasma membrane fractions revealed some physical and biochemical similarities between the light fraction of the plasma membrane and the DRM complexes, based on (1) the phosphatidylcholine/sphingomyelin ratio and (2) the protein composition visualized on a two-dimensional gel. These two fractions are also quite similar in their thermotropic phase behavior, and their high levels of ganglioside GM1. We conclude that the light membrane fraction isolated in a detergent-free environment has many of the characteristics normally associated with DRMs.

Original languageEnglish (US)
Pages (from-to)13189-13197
Number of pages9
JournalBiochemistry
Volume41
Issue number44
DOIs
StatePublished - Nov 5 2002

Fingerprint

Cell membranes
Xenopus
Detergents
Ovum
Cell Membrane
Membranes
Sphingomyelins
Cholesterol
G(M1) Ganglioside
Light
Sphingolipids
Octoxynol
Phase behavior
Phosphatidylcholines
Purification
Sucrose
Gels
Liquids
Chemical analysis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Luria, A., Vegelyte-Avery, V., Stith, B., Tsvetkova, N. M., Wolkers, W. F., Crowe, J. H., ... Nuccitelli, R. (2002). Detergent-free domain isolated from Xenopus egg plasma membrane with properties similar to those of detergent-resistant membranes. Biochemistry, 41(44), 13189-13197. https://doi.org/10.1021/bi026107b

Detergent-free domain isolated from Xenopus egg plasma membrane with properties similar to those of detergent-resistant membranes. / Luria, Ayala; Vegelyte-Avery, Vaida; Stith, Brad; Tsvetkova, Nelly M.; Wolkers, Willem F.; Crowe, John H.; Tablin, Fern; Nuccitelli, Richard.

In: Biochemistry, Vol. 41, No. 44, 05.11.2002, p. 13189-13197.

Research output: Contribution to journalArticle

Luria, A, Vegelyte-Avery, V, Stith, B, Tsvetkova, NM, Wolkers, WF, Crowe, JH, Tablin, F & Nuccitelli, R 2002, 'Detergent-free domain isolated from Xenopus egg plasma membrane with properties similar to those of detergent-resistant membranes', Biochemistry, vol. 41, no. 44, pp. 13189-13197. https://doi.org/10.1021/bi026107b
Luria, Ayala ; Vegelyte-Avery, Vaida ; Stith, Brad ; Tsvetkova, Nelly M. ; Wolkers, Willem F. ; Crowe, John H. ; Tablin, Fern ; Nuccitelli, Richard. / Detergent-free domain isolated from Xenopus egg plasma membrane with properties similar to those of detergent-resistant membranes. In: Biochemistry. 2002 ; Vol. 41, No. 44. pp. 13189-13197.
@article{4199726eccdd403b894a1d0cc945a64a,
title = "Detergent-free domain isolated from Xenopus egg plasma membrane with properties similar to those of detergent-resistant membranes",
abstract = "Microdomains known as {"}rafts{"} have been isolated from many cell types as detergent-resistant membranes (DRMs) and are enriched in sphingolipids and cholesterol. However, there has been considerable controversy over whether such domains are found in native membranes or are artificially generated by the purification procedure. This controversy is based at least in part on the fact that raft membranes were first detected following detergent extraction in the cold. We isolated two plasma membrane fractions, without detergent treatment, using a discontinuous sucrose density gradient. One fraction was designated {"}light{"} and the other {"}heavy.{"} These fractions were compared with DRMs, which were isolated in the presence of 1{\%} Triton X-100. We found that Xenopus DRMs are enriched with sphingomyelin and cholesterol and exhibit a phase state similar to the liquid-ordered phase. Comparison of DRM complexes with the light and heavy plasma membrane fractions revealed some physical and biochemical similarities between the light fraction of the plasma membrane and the DRM complexes, based on (1) the phosphatidylcholine/sphingomyelin ratio and (2) the protein composition visualized on a two-dimensional gel. These two fractions are also quite similar in their thermotropic phase behavior, and their high levels of ganglioside GM1. We conclude that the light membrane fraction isolated in a detergent-free environment has many of the characteristics normally associated with DRMs.",
author = "Ayala Luria and Vaida Vegelyte-Avery and Brad Stith and Tsvetkova, {Nelly M.} and Wolkers, {Willem F.} and Crowe, {John H.} and Fern Tablin and Richard Nuccitelli",
year = "2002",
month = "11",
day = "5",
doi = "10.1021/bi026107b",
language = "English (US)",
volume = "41",
pages = "13189--13197",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "44",

}

TY - JOUR

T1 - Detergent-free domain isolated from Xenopus egg plasma membrane with properties similar to those of detergent-resistant membranes

AU - Luria, Ayala

AU - Vegelyte-Avery, Vaida

AU - Stith, Brad

AU - Tsvetkova, Nelly M.

AU - Wolkers, Willem F.

AU - Crowe, John H.

AU - Tablin, Fern

AU - Nuccitelli, Richard

PY - 2002/11/5

Y1 - 2002/11/5

N2 - Microdomains known as "rafts" have been isolated from many cell types as detergent-resistant membranes (DRMs) and are enriched in sphingolipids and cholesterol. However, there has been considerable controversy over whether such domains are found in native membranes or are artificially generated by the purification procedure. This controversy is based at least in part on the fact that raft membranes were first detected following detergent extraction in the cold. We isolated two plasma membrane fractions, without detergent treatment, using a discontinuous sucrose density gradient. One fraction was designated "light" and the other "heavy." These fractions were compared with DRMs, which were isolated in the presence of 1% Triton X-100. We found that Xenopus DRMs are enriched with sphingomyelin and cholesterol and exhibit a phase state similar to the liquid-ordered phase. Comparison of DRM complexes with the light and heavy plasma membrane fractions revealed some physical and biochemical similarities between the light fraction of the plasma membrane and the DRM complexes, based on (1) the phosphatidylcholine/sphingomyelin ratio and (2) the protein composition visualized on a two-dimensional gel. These two fractions are also quite similar in their thermotropic phase behavior, and their high levels of ganglioside GM1. We conclude that the light membrane fraction isolated in a detergent-free environment has many of the characteristics normally associated with DRMs.

AB - Microdomains known as "rafts" have been isolated from many cell types as detergent-resistant membranes (DRMs) and are enriched in sphingolipids and cholesterol. However, there has been considerable controversy over whether such domains are found in native membranes or are artificially generated by the purification procedure. This controversy is based at least in part on the fact that raft membranes were first detected following detergent extraction in the cold. We isolated two plasma membrane fractions, without detergent treatment, using a discontinuous sucrose density gradient. One fraction was designated "light" and the other "heavy." These fractions were compared with DRMs, which were isolated in the presence of 1% Triton X-100. We found that Xenopus DRMs are enriched with sphingomyelin and cholesterol and exhibit a phase state similar to the liquid-ordered phase. Comparison of DRM complexes with the light and heavy plasma membrane fractions revealed some physical and biochemical similarities between the light fraction of the plasma membrane and the DRM complexes, based on (1) the phosphatidylcholine/sphingomyelin ratio and (2) the protein composition visualized on a two-dimensional gel. These two fractions are also quite similar in their thermotropic phase behavior, and their high levels of ganglioside GM1. We conclude that the light membrane fraction isolated in a detergent-free environment has many of the characteristics normally associated with DRMs.

UR - http://www.scopus.com/inward/record.url?scp=0037027324&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037027324&partnerID=8YFLogxK

U2 - 10.1021/bi026107b

DO - 10.1021/bi026107b

M3 - Article

C2 - 12403620

AN - SCOPUS:0037027324

VL - 41

SP - 13189

EP - 13197

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 44

ER -