Cloning and characterization of tissue inhibitor of metalloproteinase-3 (TIMP-3) from shark, Scyliorhinus torazame

We cloned the full-length cDNA encoding TIMP-3 from the cartilage of cloudy dogfish, Scyliorhinus torazame. The entire open reading frame was composed of 645 nucleotides and 214 residues including 12 conserved cysteines and a sparagine-184, a putative site for N-linked sugars. It showed about 72% ide ntity to those of other species based on the deduced amino acid sequence. T he mRNA of shark TIMP-3 were expressed abundantly in brain and cartilage ti ssues. To investigate the roles of shark TIMP-3, an expression vector was c onstructed and transfected into HT1080 human fibrosarcoma cells. Overexpres sion of shark TIMP-3 reduced the activity of MMP-2 in gelatin zymography. T hrough human Alu PCR based CAM assay, we also confirmed that shark TIMP-3 t ransfected HT1080 cells had less intravasation effects. (C) 2001 Elsevier S cience B.V. All rights reserved.