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© The Rockefeller University Press, 0021-9525/2000//1143 $5.00
The Journal of Cell Biology, Volume 149, Number 5, , 2000 1143-1156

Original Article

The Cysteine-Rich Domain of Human Adam 12 Supports Cell Adhesion through Syndecans and Triggers Signaling Events That Lead to β1 Integrin–Dependent Cell Spreading



Kousuke Ibaa, Reidar Albrechtsena, Brent Gilpina, Camilla Fröhlicha, Frosty Loechela, Anna Zolkiewskab, Kazuhiro Ishiguroc, Tetsuhito Kojimac, Wei Liud, J. Kevin Langfordd, Ralph D. Sandersond, Cord Brakebusche, Reinhard Fässlere, and Ulla M. Wewera

a The Institute of Molecular Pathology, University of Copenhagen, 2100 Copenhagen, Denmark
b Department of Biochemistry, Kansas State University, Manhattan, Kansas 66506
c Department of Medical Technology, School of Health Sciences, and First Department of Internal Medicine, Nagoya University, Nagoya 466-8550, Japan
d The Department of Pathology, Arkansas Cancer Research Center, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205
e Department of Experimental Pathology, Lund University, Lund 22185, Sweden
Institute of Molecular Pathology, University of Copenhagen, Frederik V's vej 11, 2100 Copenhagen, Denmark.45 3532 608145 3532 6056

molera{at}inet.uni2.dk

The ADAMs (a disintegrin and metalloprotease) family of proteins is involved in a variety of cellular interactions, including cell adhesion and ecto- domain shedding. Here we show that ADAM 12 binds to cell surface syndecans. Three forms of recombinant ADAM 12 were used in these experiments: the cys-teine-rich domain made in Escherichia coli (rADAM 12-cys), the disintegrin-like and cysteine-rich domain made in insect cells (rADAM 12-DC), and full-length human ADAM 12-S tagged with green fluorescent protein made in mammalian cells (rADAM 12-GFP). Mesenchymal cells specifically and in a dose-dependent manner attach to ADAM 12 via members of the syndecan family. After binding to syndecans, mesenchymal cells spread and form focal adhesions and actin stress fibers. Integrin β1 was responsible for cell spreading because function-blocking monoclonal antibodies completely inhibited cell spreading, and chondroblasts lacking β1 integrin attached but did not spread. These data suggest that mesenchymal cells use syndecans as the initial receptor for the ADAM 12 cysteine-rich domain–mediated cell adhesion, and then the β1 integrin to induce cell spreading. Interestingly, carcinoma cells attached but did not spread on ADAM 12. However, spreading could be efficiently induced by the addition of either 1 mM Mn2+ or the β1 integrin–activating monoclonal antibody 12G10, suggesting that in these carcinoma cells, the ADAM 12–syndecan complex fails to modulate the function of β1 integrin.

Key Words: ADAM • syndecan • integrin • cell adhesion • receptor cross-talk

© 2000 The Rockefeller University Press

Abbreviations used in this paper: ADAM, a disintegrin and metalloprotease; GFP, green fluorecent protein; rADAM 12-cys, recombinant cysteine-rich domain of human ADAM 12; rADAM 12-DC, recombinant disintegrin-like and cysteine-rich domain of mouse ADAM 12; rADAM 12-GFP, recombinant full-length ADAM 12-S tagged with GFP; RT, reverse transcription; SVMP, snake venom metalloprotease; TDM, triple deletion mutant.


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