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PMID:11294831

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Citation

de Marco, MC, Kremer, L, Albar, JP, Martinez-Menarguez, JA, Ballesta, J, Garcia-Lopez, MA, Marazuela, M, Puertollano, R and Alonso, MA (2001) BENE, a novel raft-associated protein of the MAL proteolipid family, interacts with caveolin-1 in human endothelial-like ECV304 cells. J. Biol. Chem. 276:23009-17

Abstract

The MAL proteolipid, an integral protein present in glycolipid- and cholesterol-enriched membrane (GEM) rafts, is an element of the machinery necessary for apical sorting in polarized epithelial Madin-Darby canine kidney cells. MAL was the first member identified of an extended family of proteins that have significant overall sequence identity. In this study we have used a newly generated monoclonal antibody to investigate an unedited member of this family, named BENE, which was found to be expressed in endothelial-like ECV304 cells and normal human endothelium. Human BENE was characterized as a proteolipid protein predominantly present in GEM rafts in ECV304 cells. Coimmunoprecipitation experiments revealed that BENE interacted with caveolin-1. Confocal immunofluorescence and electron microscopic analyses indicated that BENE mainly accumulated into intracellular vesicular/tubular structures that partially colocalize with internal caveolin-1. In response to cell surface cholesterol oxidation, BENE redistributed to the dilated vesicular structures that concentrate most of the caveolin-1 originally on the cell surface. After cessation of cholesterol oxidation, a detectable fraction of the BENE molecules migrated to the plasmalemma accompanying caveolin-1 and then returned progressively to its steady state distribution. Together, these features highlight the BENE proteolipid as being an element of the machinery for raft-mediated trafficking in endothelial cells.

Links

PubMed Online version:10.1074/jbc.M009739200

Keywords

Amino Acid Sequence; Animals; Antibodies, Monoclonal/immunology; Carrier Proteins/chemistry; Carrier Proteins/genetics; Carrier Proteins/immunology; Carrier Proteins/metabolism; Cattle; Caveolin 1; Caveolins/metabolism; Cell Line; Cholesterol/metabolism; Endothelium/cytology; Endothelium/metabolism; Endothelium/ultrastructure; Gene Expression Regulation; Humans; Membrane Proteins/chemistry; Membrane Proteins/genetics; Membrane Proteins/immunology; Membrane Proteins/metabolism; Microscopy, Immunoelectron; Molecular Sequence Data; Oxidation-Reduction; Proteolipids/chemistry; Proteolipids/metabolism; Sequence Homology, Amino Acid

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:CAV2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q13021

F

Seeded From UniProt

complete

HUMAN:CAV2

located_in

GO:0045121: membrane raft

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CAV1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q13021

F

Seeded From UniProt

complete

HUMAN:CAV1

located_in

GO:0000139: Golgi membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CAV1

located_in

GO:0005783: endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CAV1

located_in

GO:0045121: membrane raft

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MALL

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P51636

F

Seeded From UniProt

complete

HUMAN:MALL

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q03135

F

Seeded From UniProt

complete

HUMAN:MALL

located_in

GO:0000139: Golgi membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MALL

located_in

GO:0031410: cytoplasmic vesicle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MALL

located_in

GO:0045121: membrane raft

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MALL

located_in

GO:0030136: clathrin-coated vesicle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MALL

located_in

GO:0005886: plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MALL

involved_in

GO:0042632: cholesterol homeostasis

ECO:0000303: author statement without traceable support used in manual assertion

P

Seeded From UniProt

complete

HUMAN:CAV1

part_of

GO:0045121: membrane raft

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CAV1

part_of

GO:0005783: endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CAV1

involved_in

GO:0042632: cholesterol homeostasis

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

HUMAN:CAV1

involved_in

GO:0030301: cholesterol transport

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

HUMAN:CAV1

enables

GO:0015485: cholesterol binding

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

HUMAN:CAV1

part_of

GO:0000139: Golgi membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CAV2

part_of

GO:0045121: membrane raft

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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