Cholesterol substitution increases the structural heterogeneity of caveolae

Maurice Jansen, Vilja Pietiäinen, Harri Pölönen, Laura Rasilainen, Mirkka Koivusalo, Ulla Ruotsalainen, Eija Jokitalo, Elina Ikonen

Tutkimustuotos: ArtikkelijulkaisuArtikkeliTieteellinenvertaisarvioitu

Kuvaus

Caveolin-1 binds cholesterol and caveola formation involves caveolin-1 oligomerization and cholesterol association. The role of cholesterol in caveolae has so far been addressed by methods that compromise membrane integrity and abolish caveolar invaginations. To study the importance of sterol specificity for the structure and function of caveolae, we replaced cholesterol in mammalian cells with its immediate precursor desmosterol by inhibiting 24-dehydrocholesterol reductase. Desmosterol could substitute for cholesterol in maintaining cell growth, membrane integrity, and preserving caveolar invaginations. However, in desmosterol cells the affinity of caveolin-1 for sterol and the stability of caveolin oligomers were decreased. Moreover, caveolar invaginations became more heterogeneous in dimensions and in the number of caveolin-1 molecules per caveola. Despite the altered caveolar structure, caveolar ligand uptake was only moderately inhibited. We found that in desmosterol cells, Src kinase phosphorylated Cav1 at Tyr14 more avidly than in cholesterol cells. Taken the role of Cav1 Tyr14 phosphorylation in caveolar endocytosis, this may help to preserve caveolar uptake in desmosterol cells. We conclude that a sterol C24 double bond interferes with caveolin-sterol interaction and perturbs caveolar morphology but facilitates Cav1 Src phosphorylation and allows caveolar endocytosis. More generally, substitution of cholesterol by a structurally closely related sterol provides a method to selectively modify membrane protein-sterol affinity, structure and function of cholesterol-dependent domains without compromising membrane integrity.
Alkuperäiskielienglanti
LehtiJournal of Biological Chemistry
Vuosikerta283
Numero21
Sivut14610-14618
Sivumäärä9
ISSN0021-9258
DOI - pysyväislinkit
TilaJulkaistu - 2008
OKM-julkaisutyyppiA1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä, vertaisarvioitu

Lainaa tätä

Jansen, Maurice ; Pietiäinen, Vilja ; Pölönen, Harri ; Rasilainen, Laura ; Koivusalo, Mirkka ; Ruotsalainen, Ulla ; Jokitalo, Eija ; Ikonen, Elina. / Cholesterol substitution increases the structural heterogeneity of caveolae. Julkaisussa: Journal of Biological Chemistry. 2008 ; Vuosikerta 283, Nro 21. Sivut 14610-14618.
@article{a0f2f08a4f644e91a6b58aaa2c1b602b,
title = "Cholesterol substitution increases the structural heterogeneity of caveolae",
abstract = "Caveolin-1 binds cholesterol and caveola formation involves caveolin-1 oligomerization and cholesterol association. The role of cholesterol in caveolae has so far been addressed by methods that compromise membrane integrity and abolish caveolar invaginations. To study the importance of sterol specificity for the structure and function of caveolae, we replaced cholesterol in mammalian cells with its immediate precursor desmosterol by inhibiting 24-dehydrocholesterol reductase. Desmosterol could substitute for cholesterol in maintaining cell growth, membrane integrity, and preserving caveolar invaginations. However, in desmosterol cells the affinity of caveolin-1 for sterol and the stability of caveolin oligomers were decreased. Moreover, caveolar invaginations became more heterogeneous in dimensions and in the number of caveolin-1 molecules per caveola. Despite the altered caveolar structure, caveolar ligand uptake was only moderately inhibited. We found that in desmosterol cells, Src kinase phosphorylated Cav1 at Tyr14 more avidly than in cholesterol cells. Taken the role of Cav1 Tyr14 phosphorylation in caveolar endocytosis, this may help to preserve caveolar uptake in desmosterol cells. We conclude that a sterol C24 double bond interferes with caveolin-sterol interaction and perturbs caveolar morphology but facilitates Cav1 Src phosphorylation and allows caveolar endocytosis. More generally, substitution of cholesterol by a structurally closely related sterol provides a method to selectively modify membrane protein-sterol affinity, structure and function of cholesterol-dependent domains without compromising membrane integrity.",
author = "Maurice Jansen and Vilja Pieti{\"a}inen and Harri P{\"o}l{\"o}nen and Laura Rasilainen and Mirkka Koivusalo and Ulla Ruotsalainen and Eija Jokitalo and Elina Ikonen",
year = "2008",
doi = "10.1074/jbc.M710355200",
language = "English",
volume = "283",
pages = "14610--14618",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology",
number = "21",

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Cholesterol substitution increases the structural heterogeneity of caveolae. / Jansen, Maurice; Pietiäinen, Vilja; Pölönen, Harri; Rasilainen, Laura; Koivusalo, Mirkka; Ruotsalainen, Ulla; Jokitalo, Eija; Ikonen, Elina.

julkaisussa: Journal of Biological Chemistry, Vuosikerta 283, Nro 21, 2008, s. 14610-14618.

Tutkimustuotos: ArtikkelijulkaisuArtikkeliTieteellinenvertaisarvioitu

TY - JOUR

T1 - Cholesterol substitution increases the structural heterogeneity of caveolae

AU - Jansen, Maurice

AU - Pietiäinen, Vilja

AU - Pölönen, Harri

AU - Rasilainen, Laura

AU - Koivusalo, Mirkka

AU - Ruotsalainen, Ulla

AU - Jokitalo, Eija

AU - Ikonen, Elina

PY - 2008

Y1 - 2008

N2 - Caveolin-1 binds cholesterol and caveola formation involves caveolin-1 oligomerization and cholesterol association. The role of cholesterol in caveolae has so far been addressed by methods that compromise membrane integrity and abolish caveolar invaginations. To study the importance of sterol specificity for the structure and function of caveolae, we replaced cholesterol in mammalian cells with its immediate precursor desmosterol by inhibiting 24-dehydrocholesterol reductase. Desmosterol could substitute for cholesterol in maintaining cell growth, membrane integrity, and preserving caveolar invaginations. However, in desmosterol cells the affinity of caveolin-1 for sterol and the stability of caveolin oligomers were decreased. Moreover, caveolar invaginations became more heterogeneous in dimensions and in the number of caveolin-1 molecules per caveola. Despite the altered caveolar structure, caveolar ligand uptake was only moderately inhibited. We found that in desmosterol cells, Src kinase phosphorylated Cav1 at Tyr14 more avidly than in cholesterol cells. Taken the role of Cav1 Tyr14 phosphorylation in caveolar endocytosis, this may help to preserve caveolar uptake in desmosterol cells. We conclude that a sterol C24 double bond interferes with caveolin-sterol interaction and perturbs caveolar morphology but facilitates Cav1 Src phosphorylation and allows caveolar endocytosis. More generally, substitution of cholesterol by a structurally closely related sterol provides a method to selectively modify membrane protein-sterol affinity, structure and function of cholesterol-dependent domains without compromising membrane integrity.

AB - Caveolin-1 binds cholesterol and caveola formation involves caveolin-1 oligomerization and cholesterol association. The role of cholesterol in caveolae has so far been addressed by methods that compromise membrane integrity and abolish caveolar invaginations. To study the importance of sterol specificity for the structure and function of caveolae, we replaced cholesterol in mammalian cells with its immediate precursor desmosterol by inhibiting 24-dehydrocholesterol reductase. Desmosterol could substitute for cholesterol in maintaining cell growth, membrane integrity, and preserving caveolar invaginations. However, in desmosterol cells the affinity of caveolin-1 for sterol and the stability of caveolin oligomers were decreased. Moreover, caveolar invaginations became more heterogeneous in dimensions and in the number of caveolin-1 molecules per caveola. Despite the altered caveolar structure, caveolar ligand uptake was only moderately inhibited. We found that in desmosterol cells, Src kinase phosphorylated Cav1 at Tyr14 more avidly than in cholesterol cells. Taken the role of Cav1 Tyr14 phosphorylation in caveolar endocytosis, this may help to preserve caveolar uptake in desmosterol cells. We conclude that a sterol C24 double bond interferes with caveolin-sterol interaction and perturbs caveolar morphology but facilitates Cav1 Src phosphorylation and allows caveolar endocytosis. More generally, substitution of cholesterol by a structurally closely related sterol provides a method to selectively modify membrane protein-sterol affinity, structure and function of cholesterol-dependent domains without compromising membrane integrity.

U2 - 10.1074/jbc.M710355200

DO - 10.1074/jbc.M710355200

M3 - Article

VL - 283

SP - 14610

EP - 14618

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 21

ER -