Species-Specific Aggregation Factor in Sponges

An aggregation factor (AF) from the siliceous sponge Suberites domuncula has been isolated and purified by the following steps: Sepharose 2 B gel chromatography, sucrose gradient, Nonidet treatment, Sephadex G-100 gel chromatography and DEAE-Sephadex ion-exchange chromatography. By this procedure the AF was purified 1340-fold with a 63% yield nearly to homogeneity. The AF is originally associated with large particles, characterized by a sedimentation of 2200 S. These particles have been visualized electron microscopically; they are characterized by a filament-like shape of a length of 3400 Å and a cross-sectional diameter of 230 Å.
The purified, low-molecular weight AF has a buoyant density of 1.38 g/cm³ and an absorbance maximum at 282 nm. The isoelectric pH is approximately 5.75. The molecular weight of the AF has been determined to be 55,000. Chemical analysis revealed that AF consists mainly of protein.
The reaggregation process of Suberites cells to large aggregates (>1000 μm), mediated by Suberites AF, is strongly dependent on pH, ionic strength and the presence of calcium ions, and independent of incubation temperature between 0°C and 20°C. Aggregation can be inhibited by trypsin, D-glucosamine and dodecyl sulphate.
The Suberites AF is species-specific if tested in a system containing cells from the siliceous sponge Geodia cydonium .