title: Identification and characterization of an amphioxus matrix metalloproteinase homolog BbMMPL2 responding to bacteria challenge.
abstract: Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases mainly involved in extracellular matrix (ECM) degradation. We have cloned and identified BbMMPL2 as homolog of MMPs from adult amphioxus. Recombinant BbMMPL2 proteins underwent self-processing during refolding in vitro. The final ~23 kDa polypeptide displayed proteolytic activity against ECM components like casein, gelatin, collagen IV and fibrinogen, but not laminin, fibronectin or α1-PI. This activity could be inhibited by GM6001 and TIMP-1/2. In addition, real-time RT-PCR analysis revealed that BbMMPL2 expressed in all issues/organs in adult amphioxus we tested. Its transcription was significantly up-regulated 12 h post immune challenge by Escherichia coli in epidermis and hepatic diverticulum but only slightly increased by Staphyloccocus aureus in epidermis. Furthermore, recombinant BbMMPL2-EGFP expressed in 293T and NIH/3T3 cells showed aggregation in cytoplasm and induced cell death. Our results provided new evidence that MMP was involved in immune response which could be conserved through evolution.
authors.affiliation: Marine Biotechnology Research Center, School of Life Sciences, Shandong University, Jinan 250100, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Haiqing
authors.initials: H
authors.lastname: Zhang
authors.forename: Yu
authors.initials: Y
authors.lastname: Kong
authors.forename: Lijun
authors.initials: L
authors.lastname: Feng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002816:Chordata, Nonvertebrate / Q000201:enzymology / Q000235:genetics / Q000276:immunology* / Q000382:microbiology*; D004926:Escherichia coli; D020869:Gene Expression Profiling; D006801:Humans; D020782:Matrix Metalloproteinases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014162:Transfection
chemical_list: D011994:Recombinant Proteins; D020782:Matrix Metalloproteinases