Spatial segregation of transport and signalling functions between human endothelial caveolae and lipid raft proteomes

Richard R Sprenger, Ruud D Fontijn, Jan van Marle, Hans Pannekoek, Anton J G Horrevoets

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Udgivelsesdato: 2006-Dec-15
OriginalsprogEngelsk
TidsskriftBiochemical Journal
Vol/bind400
Udgave nummer3
Sider (fra-til)401-10
Antal sider9
ISSN0264-6021
DOI
StatusUdgivet - 15. dec. 2006
Udgivet eksterntJa

Fingeraftryk

Caveolae
Proteome
Lipids
Proteins
Detergents
Cell membranes
Membranes
Cell Membrane
Endothelial cells
Brain
Biotinylation
Caveolin 1
Chromosomes, Human, Pair 8
Glycosylphosphatidylinositols
Acids
Human Umbilical Vein Endothelial Cells
Vimentin
Confocal microscopy
Nitric Oxide Synthase Type III
Immunoblotting

Citer dette

Sprenger, Richard R ; Fontijn, Ruud D ; van Marle, Jan ; Pannekoek, Hans ; Horrevoets, Anton J G. / Spatial segregation of transport and signalling functions between human endothelial caveolae and lipid raft proteomes. I: Biochemical Journal. 2006 ; Bind 400, Nr. 3. s. 401-10.
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title = "Spatial segregation of transport and signalling functions between human endothelial caveolae and lipid raft proteomes",
abstract = "Lipid rafts and caveolae are biochemically similar, specialized domains of the PM (plasma membrane) that cluster specific proteins. However, they are morphologically distinct, implying different, possibly complementary functions. Two-dimensional gel electrophoresis preceding identification of proteins by MS was used to compare the relative abundance of proteins in DRMs (detergent-resistant membranes) isolated from HUVEC (human umbilical-vein endothelial cells), and caveolae immunopurified from DRM fractions. Various signalling and transport proteins were identified and additional cell-surface biotinylation revealed the majority to be exposed, demonstrating their presence at the PM. In resting endothelial cells, the scaffold of immunoisolated caveolae consists of only few resident proteins, related to structure [CAV1 (caveolin-1), vimentin] and transport (V-ATPase), as well as the GPI (glycosylphosphatidylinositol)-linked, surface-exposed protein CD59. Further quantitative characterization by immunoblotting and confocal microscopy of well-known [eNOS (endothelial nitric oxide synthase) and CAV1], less known [SNAP-23 (23 kDa synaptosome-associated protein) and BASP1 (brain acid soluble protein 1)] and novel [C8ORF2 (chromosome 8 open reading frame 2)] proteins showed different subcellular distributions with none of these proteins being exclusive to either caveolae or DRM. However, the DRM-associated fraction of the novel protein C8ORF2 (approximately 5{\%} of total protein) associated with immunoseparated caveolae, in contrast with the raft protein SNAP-23. The segregation of caveolae from lipid rafts was visually confirmed in proliferating cells, where CAV1 was spatially separated from eNOS, SNAP-23 and BASP1. These results provide direct evidence for the previously suggested segregation of transport and signalling functions between specialized domains of the endothelial plasma membrane.",
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year = "2006",
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Spatial segregation of transport and signalling functions between human endothelial caveolae and lipid raft proteomes. / Sprenger, Richard R; Fontijn, Ruud D; van Marle, Jan; Pannekoek, Hans; Horrevoets, Anton J G.

I: Biochemical Journal, Bind 400, Nr. 3, 15.12.2006, s. 401-10.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Spatial segregation of transport and signalling functions between human endothelial caveolae and lipid raft proteomes

AU - Sprenger, Richard R

AU - Fontijn, Ruud D

AU - van Marle, Jan

AU - Pannekoek, Hans

AU - Horrevoets, Anton J G

PY - 2006/12/15

Y1 - 2006/12/15

N2 - Lipid rafts and caveolae are biochemically similar, specialized domains of the PM (plasma membrane) that cluster specific proteins. However, they are morphologically distinct, implying different, possibly complementary functions. Two-dimensional gel electrophoresis preceding identification of proteins by MS was used to compare the relative abundance of proteins in DRMs (detergent-resistant membranes) isolated from HUVEC (human umbilical-vein endothelial cells), and caveolae immunopurified from DRM fractions. Various signalling and transport proteins were identified and additional cell-surface biotinylation revealed the majority to be exposed, demonstrating their presence at the PM. In resting endothelial cells, the scaffold of immunoisolated caveolae consists of only few resident proteins, related to structure [CAV1 (caveolin-1), vimentin] and transport (V-ATPase), as well as the GPI (glycosylphosphatidylinositol)-linked, surface-exposed protein CD59. Further quantitative characterization by immunoblotting and confocal microscopy of well-known [eNOS (endothelial nitric oxide synthase) and CAV1], less known [SNAP-23 (23 kDa synaptosome-associated protein) and BASP1 (brain acid soluble protein 1)] and novel [C8ORF2 (chromosome 8 open reading frame 2)] proteins showed different subcellular distributions with none of these proteins being exclusive to either caveolae or DRM. However, the DRM-associated fraction of the novel protein C8ORF2 (approximately 5% of total protein) associated with immunoseparated caveolae, in contrast with the raft protein SNAP-23. The segregation of caveolae from lipid rafts was visually confirmed in proliferating cells, where CAV1 was spatially separated from eNOS, SNAP-23 and BASP1. These results provide direct evidence for the previously suggested segregation of transport and signalling functions between specialized domains of the endothelial plasma membrane.

AB - Lipid rafts and caveolae are biochemically similar, specialized domains of the PM (plasma membrane) that cluster specific proteins. However, they are morphologically distinct, implying different, possibly complementary functions. Two-dimensional gel electrophoresis preceding identification of proteins by MS was used to compare the relative abundance of proteins in DRMs (detergent-resistant membranes) isolated from HUVEC (human umbilical-vein endothelial cells), and caveolae immunopurified from DRM fractions. Various signalling and transport proteins were identified and additional cell-surface biotinylation revealed the majority to be exposed, demonstrating their presence at the PM. In resting endothelial cells, the scaffold of immunoisolated caveolae consists of only few resident proteins, related to structure [CAV1 (caveolin-1), vimentin] and transport (V-ATPase), as well as the GPI (glycosylphosphatidylinositol)-linked, surface-exposed protein CD59. Further quantitative characterization by immunoblotting and confocal microscopy of well-known [eNOS (endothelial nitric oxide synthase) and CAV1], less known [SNAP-23 (23 kDa synaptosome-associated protein) and BASP1 (brain acid soluble protein 1)] and novel [C8ORF2 (chromosome 8 open reading frame 2)] proteins showed different subcellular distributions with none of these proteins being exclusive to either caveolae or DRM. However, the DRM-associated fraction of the novel protein C8ORF2 (approximately 5% of total protein) associated with immunoseparated caveolae, in contrast with the raft protein SNAP-23. The segregation of caveolae from lipid rafts was visually confirmed in proliferating cells, where CAV1 was spatially separated from eNOS, SNAP-23 and BASP1. These results provide direct evidence for the previously suggested segregation of transport and signalling functions between specialized domains of the endothelial plasma membrane.

U2 - 10.1042/BJ20060355

DO - 10.1042/BJ20060355

M3 - Journal article

C2 - 16886909

VL - 400

SP - 401

EP - 410

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 3

ER -