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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
21
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pubmed:dateCreated |
2004-5-17
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pubmed:abstractText |
Variant late infantile neuronal ceroid lipofuscinosis, a lysosomal storage disorder characterized by progressive mental deterioration and blindness, is caused by mutations in a polytopic membrane protein (CLN6) with unknown intracellular localization and function. In this study, transient transfection of BHK21 cells with CLN6 cDNA and immunoblot analysis using peptide-specific CLN6 antibodies demonstrated the expression of a approximately 27-kDa protein that does not undergo proteolytic processing. Cross-linking experiments revealed the presence of CLN6 dimers. Using double immunofluorescence microscopy, epitope-tagged CLN6 was shown to be retained in the endoplasmic reticulum (ER) with no colocalization with the cis-Golgi or lysosomal markers. The translocation into the ER and proper folding were confirmed by the N-linked glycosylation of a mutant CLN6 polypeptide. Pulse-chase labeling of fibroblasts from CLN6 patients and from sheep (OCL6) and mouse (nclf) models of the disease followed by immunoprecipitation of cathepsin D indicated that neither the synthesis, sorting nor the proteolytic processing of this lysosomal enzyme was affected in CLN6-defective cells. However, the degradation of the endocytosed index protein arylsulfatase A was strongly reduced in all of the mutant CLN6 cell lines compared with controls. These data suggest that defects in the ER-resident CLN6 protein lead to lysosomal dysfunctions, which may result in lysosomal accumulation of storage material.
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pubmed:grant |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/CLN6 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Cathepsin D,
http://linkedlifedata.com/resource/pubmed/chemical/Cerebroside-Sulfatase,
http://linkedlifedata.com/resource/pubmed/chemical/Cln6 protein, mouse,
http://linkedlifedata.com/resource/pubmed/chemical/Cross-Linking Reagents,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Complementary,
http://linkedlifedata.com/resource/pubmed/chemical/Epitopes,
http://linkedlifedata.com/resource/pubmed/chemical/Ligands,
http://linkedlifedata.com/resource/pubmed/chemical/Membrane Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Peptides
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pubmed:status |
MEDLINE
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pubmed:month |
May
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pubmed:issn |
0021-9258
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
21
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pubmed:volume |
279
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
22347-52
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:15010453-Animals,
pubmed-meshheading:15010453-Biotinylation,
pubmed-meshheading:15010453-Blotting, Western,
pubmed-meshheading:15010453-Cathepsin D,
pubmed-meshheading:15010453-Cell Line,
pubmed-meshheading:15010453-Cell Membrane,
pubmed-meshheading:15010453-Cerebroside-Sulfatase,
pubmed-meshheading:15010453-Cloning, Molecular,
pubmed-meshheading:15010453-Cricetinae,
pubmed-meshheading:15010453-Cross-Linking Reagents,
pubmed-meshheading:15010453-DNA, Complementary,
pubmed-meshheading:15010453-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:15010453-Endocytosis,
pubmed-meshheading:15010453-Endoplasmic Reticulum,
pubmed-meshheading:15010453-Epitopes,
pubmed-meshheading:15010453-Fibroblasts,
pubmed-meshheading:15010453-Glycosylation,
pubmed-meshheading:15010453-Golgi Apparatus,
pubmed-meshheading:15010453-Humans,
pubmed-meshheading:15010453-Immunoblotting,
pubmed-meshheading:15010453-Ligands,
pubmed-meshheading:15010453-Lysosomes,
pubmed-meshheading:15010453-Membrane Proteins,
pubmed-meshheading:15010453-Mice,
pubmed-meshheading:15010453-Mice, Inbred C57BL,
pubmed-meshheading:15010453-Microscopy, Fluorescence,
pubmed-meshheading:15010453-Mutation,
pubmed-meshheading:15010453-Peptides,
pubmed-meshheading:15010453-Precipitin Tests,
pubmed-meshheading:15010453-Protein Folding,
pubmed-meshheading:15010453-Sheep,
pubmed-meshheading:15010453-Transfection
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pubmed:year |
2004
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pubmed:articleTitle |
Defective endoplasmic reticulum-resident membrane protein CLN6 affects lysosomal degradation of endocytosed arylsulfatase A.
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pubmed:affiliation |
Department of Biochemistry, Children's Hospital, University of Hamburg, D-20246 Hamburg, Germany.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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