Suppression of membrane-type 1 matrix metalloproteinase (MMP)-mediated MMP-2 activation and tumor invasion by testican 3 and its splicing variant gene product, N-Tes
| Autor: | M, Nakada, A, Yamada, T, Takino, H, Miyamori, T, Takahashi, J, Yamashita, H, Sato |
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| Rok vydání: | 2001 |
| Předmět: |
DNA
Complementary Matrix Metalloproteinases Membrane-Associated Molecular Sequence Data Down-Regulation Protein Sorting Signals Kidney Transfection Peptide Mapping Dogs Tumor Cells Cultured Animals Humans Protein Isoforms Amino Acid Sequence RNA Messenger Cloning Molecular Gene Library Enzyme Precursors Metalloendopeptidases Matrix Metalloproteinase 16 Glioma Protein Structure Tertiary Enzyme Activation Alternative Splicing Gelatinases Proteoglycans |
| Zdroj: | Cancer research. 61(24) |
| ISSN: | 0008-5472 |
| Popis: | Using expression cloning to screen a human fetal kidney cDNA library for regulator(s) of pro-matrix metalloproteinase (MMP)-2 processing mediated by membrane-type (MT) 1 MMP, we isolated a cDNA whose product interfered with pro-MMP-2 activation. It encodes the NH(2)-terminal 313-amino acid region of a calcium-binding proteoglycan, testican 3, with a 3-amino acid substitution at the COOH terminus and thus was named N-Tes. N-Tes comprises a signal peptide, a unique domain, a follistatin-like domain, and a Ca(2+)-binding domain but lacks a COOH-terminal thyroglobulin domain and two putative glycosaminoglycan attachment sites of testican 3. Pro-MMP-2 activation by MT3-MMP was also inhibited by the coexpression of N-Tes. Immunoprecipitation analysis demonstrated direct interaction of N-Tes with either MT1-MMP or MT3-MMP. Expression of testican 1 or testican 3 but not testican 2 also inhibited pro-MMP-2 activation by either MT1-MMP or MT3-MMP. Deletion and substitution of amino acids residues in N-Tes revealed that the unique NH(2)-terminal domain of N-Tes is responsible for the inhibition of pro-MMP-2 activation by MT-MMPs. Expression of N-Tes and testican 3 was detected in normal brain but down-regulated in glioma tissues. Transfection of either the N-Tes or testican 3 gene into U251 glioma cells or Madin-Darby canine kidney cells transformed by erbB2 suppressed their invasive growth in collagen gel. These results suggest that both N-Tes and testican 3 would interfere with tumor invasion by inhibiting MT-MMPs. |
| Databáze: | OpenAIRE |
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