WISP-1 binds to decorin and biglycan.
Wnt-1-induced secreted protein 1 (WISP-1) is a member of the CCN (connective tissue growth factor, Cyr61, NOV) family of growth factors. Structural and experimental evidence suggests that CCN family member activities are modulated by their interaction with sulfated glycoconjugates. To elucidate the mechanism of action for WISP-1, we characterized the specificity of its tissue and cellular interaction and identified binding factors. WISP-1 binding was restricted to the stroma of colon tumors and to cells with a fibroblastic phenotype. By using a solid phase assay, we showed that human skin fibroblast conditioned media contained WISP-1 binding factors. Competitive inhibition with different glycosaminoglycans and treatment with glycosaminoglycan lyases and proteases demonstrated that binding to the conditioned media was mediated by dermatan sulfate proteoglycans. Mass spectrometric analysis identified the isolated binding factors as decorin and biglycan. Decorin and biglycan interacted directly with WISP-1 and inhibited its binding to components in the conditioned media. Similarly, WISP-1 interaction with human skin fibroblasts was inhibited by dermatan sulfate, decorin, and biglycan or by treatment of the cell surface with dermatan sulfate-specific lyases. Together these results demonstrate that decorin and biglycan are WISP-1 binding factors that can mediate and modulate its interaction with the surface of fibroblasts. We propose that this specific interaction plays a role in the regulation of WISP-1 function.
Desnoyers
L
L
Department of Protein Chemistry, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA. desnoyer@gene.com
Arnott
D
D
Pennica
D
D
D000818:Animals; D058578:Biglycan; D001665:Binding Sites; D001667:Binding, Competitive; D055511:CCN Intercellular Signaling Proteins; D002352:Carrier Proteins / Q000378:metabolism; D002417:Cattle; D002455:Cell Division; D002460:Cell Line; D002478:Cells, Cultured; D019765:Chondroitin ABC Lyase / Q000494:pharmacology; D003110:Colonic Neoplasms / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D049832:Decapodiformes; D058575:Decorin; D003871:Dermatan Sulfate / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D016326:Extracellular Matrix Proteins; D005347:Fibroblasts / Q000378:metabolism; D006025:Glycosaminoglycans / Q000494:pharmacology; D006133:Growth Substances / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D013058:Mass Spectrometry; D051379:Mice; D008856:Microscopy, Fluorescence; D015513:Oncogene Proteins / Q000737:chemistry / Q000378:metabolism*; D010641:Phenotype; D011485:Protein Binding; D011509:Proteoglycans / Q000737:chemistry / Q000378:metabolism*; D011518:Proto-Oncogene Proteins; D051381:Rats; D012754:Sharks; D012756:Sheep; D012867:Skin / Q000378:metabolism; D013552:Swine; D014407:Tumor Cells, Cultured
C546884:BGN protein, human; C546885:Bgn protein, mouse; C546886:Bgn protein, rat; D058578:Biglycan; D055511:CCN Intercellular Signaling Proteins; C116396:CCN4 protein, human; C492067:CCN4 protein, mouse; C492066:CCN4 protein, rat; D002352:Carrier Proteins; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; C546878:DCN protein, human; C546879:Dcn protein, mouse; C546880:Dcn protein, rat; D058575:Decorin; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D006133:Growth Substances; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D015513:Oncogene Proteins; D011509:Proteoglycans; D011518:Proto-Oncogene Proteins; D003871:Dermatan Sulfate; D019765:Chondroitin ABC Lyase