The C-terminal domain V of perlecan promotes beta1 integrin-mediated cell adhesion, binds heparin, nidogen and fibulin-2 and can be modified by glycosaminoglycans.

Publisher: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies Country of Publication: England NLM ID: 0107600 Publication Model: Print Cited Medium: Print ISSN: 0014-2956 (Print) Linking ISSN: 00142956 NLM ISO Abbreviation: Eur J Biochem Subsets: MEDLINE

Publication: -2004: Oxford, UK : Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Original Publication: Berlin, New York, Springer.

Heparan Sulfate Proteoglycans*; Cell Adhesion/*physiology ; Glycosaminoglycans/*metabolism ; Heparitin Sulfate/*metabolism ; Integrin beta1/*metabolism ; Proteoglycans/*metabolism; Animals ; Calcium-Binding Proteins/metabolism ; Chromatography, Affinity ; Extracellular Matrix Proteins/metabolism ; Fibronectins/metabolism ; Glycosaminoglycans/chemistry ; Glycoside Hydrolases/metabolism ; Heparin/metabolism ; Heparitin Sulfate/chemistry ; Heparitin Sulfate/genetics ; Heparitin Sulfate/immunology ; Hexosamines/analysis ; Humans ; Immunohistochemistry ; Integrin beta1/immunology ; Kinetics ; Membrane Glycoproteins/metabolism ; Mice ; Microscopy, Electron ; Protein Binding ; Proteoglycans/chemistry ; Proteoglycans/genetics ; Proteoglycans/immunology ; Radioimmunoassay ; Recombinant Proteins/chemistry ; Recombinant Proteins/metabolism ; Tumor Cells, Cultured

Domain V of the major basement-membrane proteoglycan perlecan, a domain which consists of three laminin type G (LG) and four epidermal-growth-factor-like (EG) modules, was obtained in recombinant form by transfecting embryonic kidney cells with an episomal expression vector. A major 90-kDa fragment V was obtained together with fragments Va (74 kDa) and Vb (26 kDa) which were generated by endogenous proteolysis in front of the most C-terminal LG module. All three fragments bound to a heparin affinity column and could be displaced at a moderate (0.2 M) NaCl concentration. Rotary-shadowing electron microscopy demonstrated a three-globule structure for fragment V. Fragment V also showed a strong immunological cross-reaction with tissue-derived perlecan, indicating that it was folded into a native structure. A further, larger fragment, Vc, was apparently substituted with heparan sulphate and/or chondroitin sulphate chains and failed to bind to heparin. Fragment V but not fragment Vc promoted a distinct adhesion of several cell lines and this could be blocked by antibodies against the integrin beta1 chain. This domain may, however, represent only one of several cell-adhesive sites of perlecan. The recombinant perlecan fragment V bound in surface plasmon resonance assays to fibulin-2, laminin-nidogen complex, nidogen and two nidogen fragments. This indicated two different nidogen-binding epitopes on perlecan domain V with about a 10-fold difference in their affinities (Kd = 0.05-0.2 microM and about 2 microM). Perlecan domain V therefore seems to participate in the supramolecular assembly and cell connections of basement membranes.

0 (Calcium-Binding Proteins)
0 (Extracellular Matrix Proteins)
0 (Fibronectins)
0 (Glycosaminoglycans)
0 (Heparan Sulfate Proteoglycans)
0 (Hexosamines)
0 (Integrin beta1)
0 (Membrane Glycoproteins)
0 (Proteoglycans)
0 (Recombinant Proteins)
0 (fibulin 2)
0 (nidogen)
143972-95-6 (perlecan)
9005-49-6 (Heparin)
9050-30-0 (Heparitin Sulfate)
EC 3.2.1.- (Glycoside Hydrolases)
EC 3.2.1.- (glycanase)

Date Created: 19980207 Date Completed: 19980205 Latest Revision: 20190620

20240829

10.1111/j.1432-1033.1997.t01-1-00039.x

9431988