Collagen fibrillogenesis during sea urchin development - Retention of SURF motifs from the N-propeptide of the 2 alpha chain in mature fibrils

The sea urchin 2 alpha fibrillar collagen chain has a unique amino-propeptide structure with several repetitions of a still unknown 140-145-amino-acid, four-Cys module called SURF (for sea urchin fibrillar module). To follow the expression of the amino-propeptide of the 2 alpha chain and assign a function to this domain, we have overproduced in Escherichia coli several recombinant proteins corresponding either to the amino-propeptide or to the amino-telopeptide. Monoclonal and/or polyclonal antibodies against these recombinant proteins allowed us to observe a similar tissue distribution during the first stages of development A signal is first observed at the prism stage as intracellular spots in mesenchymal cells. In plutei, immunofluorescence staining is observed around the skeleton spicules and as a thin meshwork surrounding the mesenchymal cells. at the ultrastructural level, and using antibodies against the amino-propeptide, gold particles are observed at the surface of 25 nm thin periodic fibrils. By rotary shadowing, these fibrils show a brush-bottle aspect, exhibiting at their surface numerous periodically distributed thin rods ended by a small globule. These data indicate that the amino-propeptide is maintained during fibrillogenesis. As previously suggested, the retention of the amino-propeptide could play an important role in regulation of the fibril growth. We propose that the important region of this amino-propeptide in thr widely encountered 25-nm-diameter fibrils is the short triple-helical segment. The globular part of the amino-propeptide will not only restrict the fibril growth but also interact with other neighbouring components and playing, as suspected from our immunofluorescence studies, a function during the spiculogenesis of the sea urchin embryo.