common genetic denominators for ca++-based skeleton in metazoa role of osteoclast-stimulating factor and of carbonic anhydrase in a calcareous sponge常见遗传分母ca + +的骨架在后生动物角色osteoclast-stimulating因子和碳酸酐酶的钙质海绵.pdfVIP

Common Genetic Denominators for Ca++-Based Skeleton in Metazoa: Role of Osteoclast-Stimulating Factor and of Carbonic Anhydrase in a Calcareous Sponge ¨ 1 1,2 1 1 1 Werner E. G. Muller *, Xiaohong Wang , Vlad A. Grebenjuk , Michael Korzhev , Matthias Wiens , 1 ¨ 1 Ute Schloßmacher , Heinz C. Schroder 1 ERC Advanced Investigator Grant Research Group at Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany, 2 National Research Center for Geoanalysis, Chinese Academy of Geological Sciences, CHN-Beijing, China Abstract Calcium-based matrices serve predominantly as inorganic, hard skeletal systems in Metazoa from calcareous sponges [phylum Porifera; class Calcarea] to proto- and deuterostomian multicellular animals. The calcareous sponges form their skeletal elements, the spicules, from amorphous calcium carbonate (ACC). Treatment of spicules from Sycon raphanus with sodium hypochlorite (NaOCl) results in the disintegration of the ACC in those skeletal elements. Until now a distinct protein/ enzyme involved in ACC metabolism could not been identified in those animals. We applied the technique of phage display combinatorial libraries to identify oligopeptides that bind to NaOCl-treated spicules: those oligopeptides allowed us to detect proteins that bind to those spicules. Two molecules have been identified, the (putative) enzyme carbonic anhydrase and the (putative) osteoclast-stimulating factor (OSTF), that are involved in the catabolism of ACC. The complete cDNAs were isolated and the recombinant proteins were prepared to raise antibodies. In turn, immu