Sponge aggregation factor: identification of the specific collagen-binding site by means of a monoclonal antibody.
The aggregation factor (AF) from the sponge Geodia cydonium is known to be a complex proteinaceous particle, composed of a series of different (glyco)proteins (Mr lower than 150,000) around a 90S sunburst-like core structure. One of the low-Mr proteins is the 47-KD cell binding fragment. We describe a new monoclonal antibody (mAb), III1E6, raised against purified AF particles, which recognizes in tissue slices structures present both on the plasma membrane and in a network-like manner in the extracellular space. By applying immunoelectron microscopical, immunoblotting, and immunoaffinity chromatographical techniques, the mAb III1E6 was shown to recognize the core structure of the AF particle. Cell adhesion studies revealed that the mAb does not inhibit AF mediated cell-cell adhesion but abolishes AF-caused attachment of cells to collagen. Electron microscopic data show that III1E6 prevents association of AF particles with collagen fibrils. By applying the techniques of immunoblotting and of protein-protein recognition on the solid phase in vitro, we could formulate the following series of events: the AF particle recognizes, with its 47-KD cell binding fragment, the aggregation receptor protein in the plasma membrane and with its core structure the collagen fibrils. These fibrils interact optionally, either via the same route or via the collagen assembly factor, with an adjacent cell surface. These findings demonstrate that the AF particle is not only the key molecule for cell-cell adhesion but also a component of cell-matrix interactions.
Gramzow
M
M
Institut für Physiologische Chemie, Universität Mainz, Federal Republic of Germany.
Schröder
H C
HC
Uhlenbruck
G
G
Batel
R
R
Müller
W E
WE
D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology; D001665:Binding Sites; D015815:Cell Adhesion Molecules; D002449:Cell Aggregation / Q000187:drug effects; D002450:Cell Communication / Q000187:drug effects; D003094:Collagen / Q000378:metabolism* / Q000494:pharmacology; D011161:Porifera; D011506:Proteins / Q000378:metabolism* / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D011506:Proteins; C013065:cell aggregation factors; D003094:Collagen