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Binding of the J 1 Adhesion Molecules to Extracellular Matrix Constituents
Author(s) -
Faissner Andreas,
Kruse Jan,
Kühn, Klaus,
Schachner Melitta
Publication year - 1990
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1990.tb02350.x
Subject(s) - laminin , extracellular matrix , glycoprotein , epitope , bovine serum albumin , fibronectin , tonicity , extracellular , biophysics , chemistry , molecular mass , matrix (chemical analysis) , biochemistry , biology , chromatography , antigen , immunology , enzyme
The J1 glycoproteins can be obtained in multiple forms in the soluble fraction of developing and adult mouse brain tissue. They are recovered as two forms of apparent molecular weights of 160,000 and 180,000 (J1‐160) from adult mouse brain and as forms of apparent molecular weights of 200,000 and 220,000 (J1‐220) from developing brain. J1‐160 and J1‐220 share common epitopes but are considered as separate entities, with J1‐220 being immunochemically closely related if not identical to tenascin. Based on the observation that J1 immunoreactivity appears on basement membrane and interstitial collagens after denervation of the neuromuscular junction in adult rodents, we became interested in investigating the binding properties of J1 glycoproteins to extracellular matrix constituents in vitro. Both J1‐160 and J1‐220 bound to collagens type I–VI and IX but not to laminin, fibronectin, bovine serum albumin, or gelatin under hypotonic buffer conditions. Under isotonic buffer conditions, J1‐220 bound to all collagen types, whereas J1‐160 bound only to collagen types V and VI with values that could be examined by Scatchard analysis. Binding of J1‐220 to collagens displayed two binding constants ( K D ) between 1.5 and 4.4 X 10 ‐9 and 1.8 and 5.5 X 10 ‐8 M , respectively, under hypotonic buffer conditions and a single K D of 2.1–8.0 X 10 ‐8 M under isotonic buffer conditions. Binding of J1‐160 to collagens had an apparent K D of 1.9–8.0 X 10 ‐9 M under hypotonic buffer conditions. Under isotonic buffer conditions, binding constants of J1‐160 to collagen types V and VI were ˜ 2 × 10 –8 M. Binding of J1‐220 to collagen type I could be inhibited by J1‐220, J1‐160, and collagen type VI but not by fibronectin or gelatin. Conversely, binding of J1‐160 was inhibited by J1‐220, J1‐160, and collagen type VI (in order of decreasing efficacy of competition). J1‐160 and J1‐220 were retained on a heparin–agarose column and eluted in a salt gradient at ˜ 0.5 M NaCl. The formation of the J1–heparin complexes was inhibited 100‐fold more efficiently by heparin than by chondroitin sulfate. These experiments show that J1 glycoproteins resemble in many respects the extracellular matrix constituents fibronectin, laminin, vitronectin, and von Willebrand factor.