Identification, Purification and Molecular Characterization of Chondrosin, a New Protein with Anti-tumoral Activity from the Marine Sponge Chondrosia Reniformis Nardo 1847.

Sonia Scarfì,Marco Giovine,Daniela Fenoglio,Marina Pozzolini,Marco Bertolino,Tiziana Altosole,Caterina Oliveri,Serena Mirata,Micha Ilan,Annalisa Salis,Gianluca Damonte

doi:10.3390/md18080409

Sonia Scarfì, Marco Giovine + Show 9 more

Open Access

https://doi.org/10.3390/md18080409

Copy DOI

Journal: Marine drugs	Publication Date: Aug 2, 2020
Citations: 9	License type: CC BY 4.0

Affiliation: University of Genoa, Tel Aviv University

Abstract

Chondrosia reniformis is a common marine demosponge showing many peculiarities, lacking silica spicules and with a body entirely formed by a dense collagenous matrix. In this paper, we have described the identification of a new cytotoxic protein (chondrosin) with selective activity against specific tumor cell lines, from C. reniformis, collected from the Liguria Sea. Chondrosin was extracted and purified using a salting out approach and molecular weight size exclusion chromatography. The cytotoxic fractions were then characterized by two-dimensional gel electrophoresis and mass spectrometry analysis and matched the results with C. reniformis transcriptome database. The procedure allowed for identifying a full-length cDNA encoding for a 199-amino acids (aa) polypeptide, with a signal peptide of 21 amino acids. The mature protein has a theoretical molecular weight of 19611.12 and an IP of 5.11. Cell toxicity assays showed a selective action against some tumor cell lines (RAW 264.7 murine leukemia cells in particular). Cell death was determined by extracellular calcium intake, followed by cytoplasmic reactive oxygen species overproduction. The in silico modelling of chondrosin showed a high structural homology with the N-terminal region of the ryanodine receptor/channel and a short identity with defensin. The results are discussed suggesting a possible specific interaction of chondrosin with the Cav 1.3 ion voltage calcium channel expressed on the target cell membranes.

Highlights

The marine sponge Chondrosia reniformis Nardo 1847 is a common marine demosponge, widely distributed along all the Mediterranean Sea and East Atlantic Ocean, where it inhabits both shallow and deep-water environments [1]
Preliminary experiments were performed on the crude hydrophilic extract (CE) of C. reniformis obtained by squeezing sponge specimens, collected in the Ligurian Sea
In this work we identified a new type of cytotoxic protein from the marine sponge C. reniformis, with unique features

Summary

Introduction

The marine sponge Chondrosia reniformis Nardo 1847 is a common marine demosponge, widely distributed along all the Mediterranean Sea and East Atlantic Ocean, where it inhabits both shallow and deep-water environments [1]. Mar. Drugs 2020, 18, 409 body is entirely formed by a dense collagenous matrix with peculiar molecular features [2,3,4] with finely-regulated production [5]. Drugs 2020, 18, 409 body is entirely formed by a dense collagenous matrix with peculiar molecular features [2,3,4] with finely-regulated production [5] This sponge is characterized by good regenerative properties [6].

Objectives

Methods

Results

Conclusion