title: Endocytosis of the dermatan sulfate proteoglycan decorin utilizes multiple pathways and is modulated by epidermal growth factor receptor signaling.
abstract: Human skin fibroblasts efficiently internalize the matrikine decorin by receptor-mediated endocytosis, however, very little is known about its intracellular trafficking routes up to lysosomal degradation. In an in vitro system measuring uptake and degradation of [(35)S]sulfate-labeled decorin, endocytosis was blocked by 46% when clathrin assembly/disassembly was inhibited using chlorpromazine. Pharmacological inhibition of EGF receptor signaling caused 34% reduction of decorin uptake, whereas inhibition of the IGF receptor had no effect. Using confocal immunofluorescence microscopy, we determined that only about 5-10% of internalized decorin colocalized with the EGFR. Thus, uptake depends on EGFR signaling rather than trafficking along the same pathway. Decorin passes through early endosomes towards trafficking to lysosomes, since more than 50% of decorin colocalized with EEA1. Moreover, inhibition of endosomal fusion by wortmannin caused a profound inhibition of decorin endocytosis. Overexpression of the clathrin-binding Hrs protein, which has previously been shown to inibit EGFR degradation blocked the degradation of decorin. Cholesterol depletion by filipin inhibited uptake of decorin by 34%, however, nearly no intracellular colocalization was found between decorin and caveolin-1. The combined use of filipin and chlorpromazine had an additive inhibitory effect on decorin endocytosis. Moreover, chlorpromazine diverted decorin from the chlorpromazine-sensitive pathway to an alternative uptake route. The CD44/hyaluronan pathway was excluded as an endocytic route for decorin. Our observations indicate that decorin is taken up by more than one endocytic pathway. Of note, lipid-raft-dependent EGFR signaling modulates decorin uptake, suggesting the presence of a potential feedback regulation mechanism for desensitization of signaling events mediated by decorin.
authors.affiliation: Department of Obstetrics and Gynecology, Domagkstr. 11, Münster University Hospital, D-48149 Münster, Germany.
authors.forename: David Denis Sofeu
authors.initials: DD
authors.lastname: Feugaing
authors.forename: Raija
authors.initials: R
authors.lastname: Tammi
authors.forename: Frank G
authors.initials: FG
authors.lastname: Echtermeyer
authors.forename: Harald
authors.initials: H
authors.lastname: Stenmark
authors.forename: Hans
authors.initials: H
authors.lastname: Kresse
authors.forename: Martin
authors.initials: M
authors.lastname: Smollich
authors.forename: Elke
authors.initials: E
authors.lastname: Schönherr
authors.forename: Ludwig
authors.initials: L
authors.lastname: Kiesel
authors.forename: Martin
authors.initials: M
authors.lastname: Götte
mesh_terms: D002746:Chlorpromazine / Q000494:pharmacology; D002784:Cholesterol; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism; D002966:Clathrin; D058575:Decorin; D003871:Dermatan Sulfate / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D066246:ErbB Receptors / Q000378:metabolism / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005347:Fibroblasts; D005372:Filipin / Q000494:pharmacology; D006801:Humans; D021381:Protein Transport; D011509:Proteoglycans / Q000378:metabolism*; D015398:Signal Transduction; D012867:Skin
chemical_list: D011508:Chondroitin Sulfate Proteoglycans; D002966:Clathrin; C546878:DCN protein, human; D058575:Decorin; D016326:Extracellular Matrix Proteins; D011509:Proteoglycans; C032649:dermatan sulfate proteoglycan; D003871:Dermatan Sulfate; D005372:Filipin; D002784:Cholesterol; D066246:ErbB Receptors; D002746:Chlorpromazine