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Cited by 203

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Both Sphingolipids and Cholesterol Participate in the Detergent Insolubility of Alkaline Phosphatase, a Glycosylphosphatidylinositol-anchored Protein, in Mammalian Membranes

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Hanada, Kentaro, Nishijima, Masahiro, Akamatsu, Yuzuru, Pagano, Richard E.·1995·Journal of Biological Chemistry
DOI 10.1074/jbc.270.11.6254
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Abstract

SPB-1, a Chinese hamster ovary cell variant defective in serine palmitoyltransferase activity for sphingolipid synthesis, provides a useful system for studying the effects of sphingolipids and/or cholesterol deprivation on cellular functions and membrane properties. To investigate whether there was an interaction among sphingolipids, cholesterol, and glycosylphosphatidylinositol (GPI)-anchored proteins in biological membranes, we introduced human placental alkaline phosphatase (PLAP) in SPB-1 and in wild type cells by stable transfection and examined the effects of sphingolipid and/or cholesterol deprivation on the solubility of PLAP in Triton X-100. Although the PLAP solubility of the membranes isolated from the control cells in Triton X-100 was only 10%, deprivation of sphingolipid and cholesterol further enhanced the solubility, which reached 50% when both sphingolipids and cholesterol were deprived. The enhanced solubility was suppressed to the control level by metabolic complementation with exogenous sphingosine and cholesterol. The sphingolipid and cholesterol content of the isolated membranes changed independently, eliminating the possibility that sphingolipid deprivation induced a reduction in cellular cholesterol and enhanced PLAP solubility and vice versa. It was also unlikely that the enhanced solubility was due to structural changes in PLAP molecules since, regardless of sphingolipid and cholesterol deprivations, almost all PLAP had the GPI-anchor moiety and there were no differences in the apparent molecular weight of the protein in supernatant and precipitate fractions of the detergent-treated membranes. In addition, the expression level of caveolin in the isolated membranes was not significantly affected by sphingolipids and/or cholesterol depletion. These results indicated that both sphingolipids and cholesterol were involved in the PLAP insolubility and suggested that these lipids coordinately played a role in formation of Triton X-100-resistant complexes.

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Metadata

Authors
Hanada, K., Nishijima, M., Akamatsu, Y., & Pagano, R. E.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1995-03
Volume
270
Issue
11
Pages
6254-6260
ISSN
0021-9258 (ISSN-L)
DOI
10.1074/jbc.270.11.6254
OpenAlex
W1976010795
Cited by
203
References
2
FWCI
6.75
Type
journal article
Language
en
Open access
hybrid
Licence

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CoChat. (2026). Citation check: Both Sphingolipids and Cholesterol Participate in the Detergent Insolubility of Alkaline Phosphatase, a Glycosylphosphatidylinositol-anchored Protein, in Mammalian Membranes. https://app.cochat.ai/papers/10.1074/jbc.270.11.6254
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Hanada, K., Nishijima, M., Akamatsu, Y., & Pagano, R. E. (1995). Both Sphingolipids and Cholesterol Participate in the Detergent Insolubility of Alkaline Phosphatase, a Glycosylphosphatidylinositol-anchored Protein, in Mammalian Membranes. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.270.11.6254

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