Premium
Purification and partial characterization of Xenophus laevis tenascin from the XTC cell line
Author(s) -
Riou J.F.,
Alfandari D.,
Eppe M.,
Tacchetti C.,
Chiquet M.,
Boucaut J.C.,
Thiery J.P.,
Levi G.
Publication year - 1991
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(91)80184-5
Subject(s) - xenopus , tenascin , polyclonal antibodies , extracellular matrix , affinity chromatography , microbiology and biotechnology , monoclonal antibody , laminin , basal lamina , chemistry , biology , biochemistry , fibronectin , antibody , anatomy , ultrastructure , immunology , gene , enzyme
We report here the purification of tenascin, an extracellular matrix molecule involved in the control of morphogenesis, from the conditioned medium of the Xenopus XTC cell line. Tenascin was purified by affinity chromatography on a column of the monoclonal antibody mAb TnM1; the molecule eluted from this column has a relative molecular mass of 210 kDa after reduction. Electrophoretic analysis under non‐reducing conditions shows that the purified components are oligomeric disulfide‐linked complexes which barely enter # 4% polyacrylamide gel. Upon rotary shadowing these molecules appear to possess a central globular domain to which pairs or triplets of arms are attached. Polyclonal antibodies have been raised against purified Xenopus tenascin. They recognise specifically the antigen on Western blots of XTC conditioned medium and adult brain by immunofluorescence, these antibodies reveal large amounts of tenascin in the secretory vesicles as well as in the extracellular matrix of XTC cells. In the Xenopus tadpole, they stain the developing cartilage, the basal lumina of skin epidermis, myotendinous ligaments and restricted regions of the central nervous system.