Abstract
Nidogen-1 and nidogen-2 are homologous proteins found in all basement membranes (BMs). They show comparable binding activities in vitro and partially redundant functions in vivo.
Previously, we showed that in skin organotypic cocultures, BM formation
was prevented in the absence of nidogens and that either nidogen was
able to rescue this failure. We now dissected the two nidogens to
identify the domains required for BM deposition. For that purpose, HaCaT
cells were grown on collagen matrices containing nidogen-deficient,
murine fibroblasts. After addition of nidogen-1 or nidogen-2 protein
fragments comprising different binding domains, BM deposition was
analyzed by immunofluorescence and electron microscopy. We could
demonstrate that the rod-G3 domain of nidogen-2 was sufficient to
achieve deposition of BM components at the epidermal-collagen interface.
In contrast, for nidogen-1, both the G2 and G3 domains were required.
Immunoblot analysis confirmed that all BM components were present in
comparable amounts under all culture conditions. This finding
demonstrates that nidogens, although homologous proteins, exert their
effect on BM assembly through different binding domains, which may in
turn result in alterations of BM structure and functions, thus providing
an explanation for the phenotypical differences observed between
nidogen-1 and -2 deficient mice.—Bechtel, M., Keller, M. V., Bloch, W.,
Sasaki, T., Boukamp, P., Zaucke, F., Paulsson, M., Nischt, R. Different
domains in nidogen-1 and nidogen-2 drive basement membrane formation in
skin organotypic cocultures. FASEB J. 26, 3637–3648 (2012). www.fasebj.org
Original language | English |
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Journal | FASEB journal : official publication of the Federation of American Societies for Experimental Biology |
Volume | 26 |
Issue number | 9 |
Pages (from-to) | 3637-3648 |
Number of pages | 12 |
DOIs | |
Publication status | Published - 09.2012 |
Research areas and keywords
- Base Sequence
- Basement Membrane
- Binding Sites
- Cell Line
- Coculture Techniques
- Culture Media, Conditioned
- DNA Primers
- Fluorescent Antibody Technique, Indirect
- Laminin
- Membrane Glycoproteins
- Microscopy, Electron, Transmission
- Polymerase Chain Reaction
- Skin