Purification and partial characterization of Xenopus laevis tenascin from the XTC cell line.
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 a 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 lamina of skin epidermis, myotendinous ligaments and restricted regions of the central nervous system.
Riou
J F
JF
Laboratorie de Biologie Expérimentale, CNRS, Université P. et M. Curie, Paris, France.
Alfandari
D
D
Eppe
M
M
Tacchetti
C
C
Chiquet
M
M
Boucaut
J C
JC
Thiery
J P
JP
Levi
G
G
D000818:Animals; D015153:Blotting, Western; D015816:Cell Adhesion Molecules, Neuronal / Q000276:immunology / Q000302:isolation & purification* / Q000648:ultrastructure; D002460:Cell Line; D016326:Extracellular Matrix Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D008854:Microscopy, Electron; D008970:Molecular Weight; D019063:Tenascin; D014982:Xenopus laevis / Q000196:embryology
D015816:Cell Adhesion Molecules, Neuronal; D016326:Extracellular Matrix Proteins; D019063:Tenascin