Membrane proteins exposed on the external side of the intestinal brush-border membrane have fusogenic properties.
The intestinal brush-border membrane contains one or several membrane proteins that mediate fusion and/or aggregation of small unilamellar egg phosphatidylcholine vesicles. The fusion is accompanied by a partial loss of vesicle contents. Proteolytic treatment of the brush-border membrane with proteinase K abolishes the fusogenic property. This finding suggests that the fusogenic activity is associated with a membrane protein exposed on the external or luminal side of the brush-border membrane. Activation of intrinsic proteinases of the brush-border membrane liberates water-soluble proteins (supernate proteins). These proteins behave in an analogous way to intact brush-border membrane vesicles; they induce fusion of egg phosphatidylcholine vesicles and render the egg phosphatidylcholine bilayer permeable to ions and small molecules (Mr less than or equal to 5000). Furthermore, supernate proteins mediate phosphatidylcholine and cholesterol exchange between two populations of small, unilamellar phospholipid vesicles. Supernate proteins are fractionated on Sephadex G-75 SF yielding three protein peaks of apparent Mr greater than or equal to 70,000, Mr = 22,000 and Mr = 11,500. All three protein fractions show similar phosphatidylcholine-exchange activity, but they differ in their effects on the stability of egg phosphatidylcholine vesicles. The protein fraction with an apparent Mr greater than or equal to 70,000 has the highest fusogenic activity while the protein fraction of apparent Mr = 11,500 appears to be most effective in rendering the egg phosphatidylcholine bilayer permeable.
Laboratorium für Biochemie, Eidgenössische Technische Hochschule Zürich, ETH-Zentrum, Switzerland.
H
H
Thurnhofer
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G
Lipka
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H
Hauser
D000818:Animals; D002850:Chromatography, Gel; D019286:Endopeptidase K; D005452:Fluoresceins / Q000378:metabolism; D005614:Freeze Fracturing; D007422:Intestines / Q000648:ultrastructure*; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000502:physiology*; D008854:Microscopy, Electron; D008871:Microvilli / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000378:metabolism; D011817:Rabbits; D012697:Serine Endopeptidases / Q000494:pharmacology; D013050:Spectrometry, Fluorescence
D005452:Fluoresceins; D008051:Lipid Bilayers; D008081:Liposomes; D008565:Membrane Proteins; D010713:Phosphatidylcholines; C024098:6-carboxyfluorescein; D012697:Serine Endopeptidases; D019286:Endopeptidase K