Back to Search
Start Over
Nephronectin binds to heparan sulfate proteoglycans via its MAM domain
- Source :
- Matrix Biology. 32:188-195
- Publication Year :
- 2013
- Publisher :
- Elsevier BV, 2013.
-
Abstract
- Nephronectin is a basement membrane protein comprising five N-terminal epidermal growth factor (EGF)-like repeats, a central linker segment containing an Arg-Gly-Asp (RGD) motif and a C-terminal meprin-A5 protein-receptor protein tyrosine phosphatase μ (MAM) domain. Nephronectin has been shown to interact with α8β1 integrin through the central linker segment, but its interactions with other molecules remain to be elucidated. Here, we examined the binding of nephronectin to a panel of glycosaminoglycan (GAG) chains. Nephronectin bound strongly to heparin and chondroitin sulfate (CS)-E and moderately to heparan sulfate (HS), but failed to bind to CS-A, CS-C, CS-D, dermatan sulfate and hyaluronic acid. Deletion of the MAM domain severely impaired the binding of nephronectin to heparin but not CS-E, whereas deletion of the EGF-like repeats reduced its binding to CS-E but not heparin, suggesting that nephronectin interacts with CS-E and heparin through the EGF-like repeats and MAM domain, respectively. Consistent with these results, nephronectin bound to agrin and perlecan, which are heparan sulfate proteoglycans (HSPGs) in basement membranes, in HS-dependent manners. Site-directed mutagenesis of the MAM domain revealed that multiple basic amino acid residues in the putative loop regions were involved in the binding of the MAM domain to agrin. The binding of nephronectin to basement membrane HSPGs was further confirmed by in situ nephronectin overlay assays using mouse frozen tissue sections. Taken together, these findings indicate that nephronectin is capable of binding to HSPGs in basement membranes via the MAM domain, and thereby raise the possibility that interactions with basement membrane HSPGs may be involved in the deposition of nephronectin onto basement membranes.
- Subjects :
- Models, Molecular
Molecular Sequence Data
Perlecan
Plasma protein binding
Biology
Kidney
Basement Membrane
Protein Structure, Secondary
Dermatan sulfate
Mice
chemistry.chemical_compound
medicine
Animals
Humans
Protein Interaction Domains and Motifs
Agrin
Amino Acid Sequence
Chondroitin sulfate
Lung
Molecular Biology
Basement membrane
Extracellular Matrix Proteins
Kidney metabolism
Heparan sulfate
Embryo, Mammalian
Molecular biology
Cell biology
carbohydrates (lipids)
HEK293 Cells
medicine.anatomical_structure
chemistry
biology.protein
Heparan Sulfate Proteoglycans
Protein Binding
Subjects
Details
- ISSN :
- 0945053X
- Volume :
- 32
- Database :
- OpenAIRE
- Journal :
- Matrix Biology
- Accession number :
- edsair.doi.dedup.....2f876841ef9eba495261cd987a45cc73