Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity
| Autor: | Yoshiko Yagi, Hitomi Fujisaki, Maria Hayashi, Koji Kimata, Nobuo Sugiura, Yuya Sato, Tetsuo Okubo, Shunji Hattori, Naoko Norioka, Shaoliang Li, Kiyotoshi Sekiguchi, Itsuko Nakano, Chisei Shimono |
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| Rok vydání: | 2010 |
| Předmět: |
endocrine system
animal structures Blotting Western Molecular Sequence Data Glycobiology and Extracellular Matrices Perlecan Biochemistry Immunoenzyme Techniques Extracellular matrix Mice chemistry.chemical_compound Animals Humans Amino Acid Sequence Luciferases Molecular Biology Activin type 2 receptors Extracellular Matrix Proteins Membrane Glycoproteins Agrin Sequence Homology Amino Acid biology Heparin Cell Biology Heparan sulfate Embryo Mammalian Recombinant Proteins Activins carbohydrates (lipids) Fibronectin chemistry embryonic structures Mutagenesis Site-Directed biology.protein Heparan sulfate binding Heparitin Sulfate Laminin Heparan Sulfate Proteoglycans hormones hormone substitutes and hormone antagonists ACVR2B Protein Binding |
| Zdroj: | Journal of Biological Chemistry. 285:36645-36655 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m110.177865 |
| Popis: | This research was originally published in the Journal of Biological Chemistry. Shaoliang Li, Chisei Shimono, Naoko Norioka, Itsuko Nakano, Tetsuo Okubo, Yoshiko Yagi, Maria Hayashi, Yuya Sato, Hitomi Fujisaki, Shunji Hattori, Nobuo Sugiura, Koji Kimata and Kiyotoshi Sekiguchi. Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity. J. Biol. Chem. 2010; 285: 36645-36655 © the American Society for Biochemistry and Molecular Biology Activin A, a member of the transforming growth factor-β family, plays important roles in hormonal homeostasis and embryogenesis. In this study, we produced recombinant human activin A and examined its abilities to bind to extracellular matrix proteins. Recombinant activin A expressed in 293-F cells was purified as complexes of mature dimeric activin A with its pro-region. Among a panel of extracellular matrix proteins tested, recombinant activin A bound to perlecan and agrin, but not to laminins, nidogens, collagens I and IV, fibronectin, and nephronectin. The binding of recombinant activin A to perlecan was inhibited by heparin and high concentrations of NaCl and abolished by heparitinase treatment of perlecan, suggesting that activin A binds to the heparan sulfate chains of perlecan. In support of this possibility, recombinant activin A was capable of directly binding to heparin and heparan sulfate chains. Site-directed mutagenesis of recombinant activin A revealed that clusters of basic amino acid residues, Lys259-Lys263 and Lys270-Lys272, in the pro-region were required for binding to perlecan. Interestingly, deletion of the peptide segment Lys259-Gly277 containing both basic amino acid clusters from the pro-region did not impair the activity of activin A to stimulate Smad-dependent gene expressions, although it completely ablated the perlecan-binding activity. The binding of activin A to basement membrane heparan sulfate proteoglycans through the basic residues in the pro-region was further confirmed by in situ activin A overlay assays using frozen tissue sections. Taken together, the present results indicate that activin A binds to heparan sulfate proteoglycans through its pro-region and thereby regulates its localization within tissues. |
| Databáze: | OpenAIRE |
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