title: Comparative analysis of protein profiles of aqueous extracts from marine sponges and assessment of cytotoxicity on different mammalian cell types.
abstract: Marine natural products extracted from sponges represent a new source for drug discovery. Here we describe a simple method for preparing aqueous extracts from 7 Mediterranean demosponges, which allowed the extraction of water-soluble compounds, such as proteins by homogenization of sponge tissue in phosphate buffered saline (PBS). The comparative analysis by SDS-PAGE showed differences in number of bands, bandwidth and intensity among the sponges analyzed. The PAS/silver staining revealed a substantial and different glycoprotein assortment among the demosponges studied. To further study the biological activities present in the sponge extracts, we determined the non-cytotoxic doses on four different mammalian cell types demonstrating that the optimal non-cytotoxic doses were cell type- and extract-dependent. In conclusion, the extraction method described in this paper represents a fast and efficient procedure for the extraction of water-soluble proteins from marine sponges. Furthermore, the cell viability data suggest the feasibility of this method for the direct in vitro cell-based assays.
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Via Orabona 4, 70126 Bari, Italy.
authors.forename: Gaetano
authors.initials: G
authors.lastname: Di Bari
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Via Orabona 4, 70126 Bari, Italy.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Gentile
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Via Orabona 4, 70126 Bari, Italy.
authors.forename: Tiziana
authors.initials: T
authors.lastname: Latronico
authors.affiliation: Department of Biology, University of Bari, Via Orabona 4, 70126 Bari, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Corriero
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Via Orabona 4, 70126 Bari, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Fasano
authors.affiliation: Department of Biology, University of Bari, Via Orabona 4, 70126 Bari, Italy.
authors.forename: Carlotta Nonnis
authors.initials: CN
authors.lastname: Marzano
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Via Orabona 4, 70126 Bari, Italy. Electronic address: graziamaria.liuzzi@uniba.it.
authors.forename: Grazia Maria
authors.initials: GM
authors.lastname: Liuzzi
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000187:drug effects*; D001688:Biological Products / Q000032:analysis / Q000494:pharmacology; D018938:Caco-2 Cells; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D020714:Maximum Tolerated Dose; D011161:Porifera / Q000737:chemistry* / Q000145:classification; D011506:Proteins / Q000032:analysis* / Q000494:pharmacology; D051381:Rats; D012995:Solubility
chemical_list: D001688:Biological Products; D006023:Glycoproteins; D011506:Proteins
keywords: Aqueous extracts; Bioactive compounds; Cell morphology; Demosponges; In vitro cytotoxicity