Expression of fibulin-2 by fibroblasts and deposition with fibronectin into a fibrillar matrix.
The extracellular matrix protein fibulin-2 was shown to be a typical product of cultured human and mouse fibroblasts by several immunological assays. It is secreted and deposited in cells and tissues as a disulfide-bonded oligomer identical in size to the previously described recombinant fibulin-2. Most of the fibroblast fibulin-2 is deposited into a dense fibrillar meshwork which requires treatment with EDTA and/or 6 M urea for solubilization. Fibulin-2 and fibronectin are synthesized at equivalent levels and both colocalize in the fibrils as shown by immunofluorescence. Metabolic labelling and pulse-chase studies demonstrated fibulin-2 oligomers in detergent extracts of cells and their rapid translocation to extracellular EDTA-sensitive assembly forms. Unlike for fibronectin and fibulin-1 only a little fibulin-2 was found in the cell culture medium. Immunogold staining of confluent human fibroblasts showed localization of fibulin-2 to a fine meshwork or bundles of amorphous microfibrils in the matrix. This also demonstrated a distinct colocalization of fibulin-2 and fibronectin at the electron microscope level, indicating that the interaction between these two protein shown in in vitro assays may also exist in situ. No distinct colocalization of both proteins could, however, be observed with cross-striated fibrils of collagen I and collagen VI microfibrils.
Sasaki
T
T
Max-Planck-Institut für Biochemie, Martinsried, Germany.
Wiedemann
H
H
Matzner
M
M
Chu
M L
ML
Timpl
R
R
D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D004492:Edetic Acid / Q000494:pharmacology; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008854:Microscopy, Electron
D002135:Calcium-Binding Proteins; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; C084612:fibulin 2; D004492:Edetic Acid