title: The membrane-associated form of α(s1)-casein interacts with cholesterol-rich detergent-resistant microdomains.
abstract: Caseins, the main milk proteins, interact with colloidal calcium phosphate to form the casein micelle. The mesostructure of this supramolecular assembly markedly influences its nutritional and technological functionalities. However, its detailed molecular organization and the cellular mechanisms involved in its biogenesis have been only partially established. There is a growing body of evidence to support the concept that α(s1)-casein takes center stage in casein micelle building and transport in the secretory pathway of mammary epithelial cells. Here we have investigated the membrane-associated form of α(s1)-casein in rat mammary epithelial cells. Using metabolic labelling we show that α(s1)-casein becomes associated with membranes at the level of the endoplasmic reticulum, with no subsequent increase at the level of the Golgi apparatus. From morphological and biochemical data, it appears that caseins are in a tight relationship with membranes throughout the secretory pathway. On the other hand, we have observed that the membrane-associated form of α(s1)-casein co-purified with detergent-resistant membranes. It was poorly solubilised by Tween 20, partially insoluble in Lubrol WX, and substantially insoluble in Triton X-100. Finally, we found that cholesterol depletion results in the release of the membrane-associated form of α(s1)-casein. These experiments reveal that the insolubility of α(s1)-casein reflects its partial association with a cholesterol-rich detergent-resistant microdomain. We propose that the membrane-associated form of α(s1)-casein interacts with the lipid microdomain, or lipid raft, that forms within the membranes of the endoplasmic reticulum, for efficient forward transport and sorting in the secretory pathway of mammary epithelial cells.
authors.affiliation: UR1196 Génomique et Physiologie de la Lactation, Institut National de la Recherche Agronomique, Jouy-en-Josas, France.
authors.forename: Annabelle
authors.initials: A
authors.lastname: Le Parc
authors.affiliation: UR1196 Génomique et Physiologie de la Lactation, Institut National de la Recherche Agronomique, Jouy-en-Josas, France.
authors.forename: Edith
authors.initials: E
authors.lastname: Honvo Houéto
authors.affiliation: UR1196 Génomique et Physiologie de la Lactation, Institut National de la Recherche Agronomique, Jouy-en-Josas, France.
authors.forename: Natascha
authors.initials: N
authors.lastname: Pigat
authors.affiliation: UR1196 Génomique et Physiologie de la Lactation, Institut National de la Recherche Agronomique, Jouy-en-Josas, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Chat
authors.affiliation: UMR1253 Science et Technologie du Lait et de l'Œuf, Institut National de la Recherche Agronomique, Rennes, France.
authors.forename: Joëlle
authors.initials: J
authors.lastname: Leonil
authors.affiliation: UR1196 Génomique et Physiologie de la Lactation, Institut National de la Recherche Agronomique, Jouy-en-Josas, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Chanat
mesh_terms: D000818:Animals; D001692:Biological Transport; D002364:Caseins / Q000737:chemistry* / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry*; D003902:Detergents / Q000494:pharmacology; D004847:Epithelial Cells / Q000378:metabolism / Q000648:ultrastructure; D019143:Evolution, Molecular; D005260:Female; D007774:Lactation; D008322:Mammals / Q000378:metabolism*; D021962:Membrane Microdomains / Q000737:chemistry* / Q000187:drug effects; D008823:Micelles; D017208:Rats, Wistar; D013045:Species Specificity
chemical_list: D002364:Caseins; D003902:Detergents; D008823:Micelles; D002784:Cholesterol