title: Biochemical characterization of MLC1 protein in astrocytes and its association with the dystrophin-glycoprotein complex.
abstract: MLC1 gene mutations have been associated with megalencephalic leukoencephalopathy with subcortical cysts (MLC), a rare neurologic disorder in children. The MLC1 gene encodes a membrane protein (MLC1) with unknown function which is mainly expressed in astrocytes. Using a newly developed anti-human MLC1 polyclonal antibody, we have investigated the biochemical properties and localization of MLC1 in cultured astrocytes and brain tissue and searched for evidence of a relationship between MLC1 and proteins of the dystrophin-glycoprotein complex (DGC). Cultured astrocytes express two MLC1 components showing different solubilisation properties and subcellular distribution. Most importantly, we show that the membrane-associated component of MLC1 (60-64 kDa) localizes in astrocytic lipid rafts together with dystroglycan, syntrophin and caveolin-1, and co-fractionates with the DGC in whole rat brain tissue. In the human brain, MLC1 protein is expressed in astrocyte processes and ependymal cells, where it colocalizes with dystroglycan and syntrophin. These data indicate that the DGC may be involved in the organization and function of the MLC1 protein in astrocyte membranes.
authors.affiliation: Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. elena.ambrosini@iss.it
authors.forename: Elena
authors.initials: E
authors.lastname: Ambrosini
authors.forename: Barbara
authors.initials: B
authors.lastname: Serafini
authors.forename: Angela
authors.initials: A
authors.lastname: Lanciotti
authors.forename: Fabio
authors.initials: F
authors.lastname: Tosini
authors.forename: Flavia
authors.initials: F
authors.lastname: Scialpi
authors.forename: Rossana
authors.initials: R
authors.lastname: Psaila
authors.forename: Carla
authors.initials: C
authors.lastname: Raggi
authors.forename: Francesco
authors.initials: F
authors.lastname: Di Girolamo
authors.forename: Tamara Corinna
authors.initials: TC
authors.lastname: Petrucci
authors.forename: Francesca
authors.initials: F
authors.lastname: Aloisi
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D001254:Astrocytoma / Q000473:pathology; D001921:Brain / Q000166:cytology; D002478:Cells, Cultured; D049030:Dystroglycans / Q000378:metabolism*; D049029:Dystrophin-Associated Proteins / Q000378:metabolism; D006801:Humans; D021962:Membrane Microdomains / Q000378:metabolism; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D009363:Neoplasm Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D049029:Dystrophin-Associated Proteins; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D009363:Neoplasm Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; C091824:syntrophin; D049030:Dystroglycans