title: Distal M domain of cobra ADAM-like metalloproteinase mediates the binding of positively charged cysteine-rich domain to αvβ3 integrin in the suppression of cell migration.
abstract: We have previously identified two new P-III type ADAM-like snake venom metalloproteinases (SVMPs), i.e., atragin and kaouthiagin-like, from Taiwan cobra venom and determined their 3D structures with a distinct C- and I-shaped metalloproteinase/disintegrin-like/cysteine-rich (MDC) modular architecture. Herein, we investigated their functional targets to elucidate the role of cobra SVMPs in perturbing wound healing in snakebite victims. We showed that the non-RGD (Arg-Gly-Asp) C-shaped SVMP atragin binds about ten-fold stronger than the RGD-containing I-shaped SVMP kaouthiagin-like to αvβ3 integrin in the surface-immobilized form. Atragin binds to αvβ3 integrin through a novel interaction mode involving distal M and C domains via the RRN sequence motif in the hyper variable loop. In a cell adhesion assay, the adhesion of fibroblasts to atragin was mediated by αvβ3 integrin. Furthermore, atragin inhibited wound healing and suppressed cell migration in a αvβ3 integrin-dependent manner. These results, together with our previous demonstration of non-cytotoxic cobra CTX A5 in targeting αvβ3 integrin, suggest that cobra venom consists of several non-RGD toxins with integrin-binding specificity that could perturb wound healing in snakebite victims.
authors.affiliation: Department of Life Sciences and Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 30013, Taiwan; Refining & Manufacturing Research Institute, CPC Corporation, Chia-Yi 60051, Taiwan.
authors.forename: Po-Long
authors.initials: PL
authors.lastname: Wu
authors.affiliation: Department of Life Sciences and Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 30013, Taiwan.
authors.forename: Chien-Chu
authors.initials: CC
authors.lastname: Lin
authors.affiliation: Department of Life Sciences and Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 30013, Taiwan; National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Miaoli 35053, Taiwan.
authors.forename: Ting-Hui
authors.initials: TH
authors.lastname: Lin
authors.affiliation: National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Miaoli 35053, Taiwan.
authors.forename: Min-Shi
authors.initials: MS
authors.lastname: Lee
authors.affiliation: Department of Life Sciences and Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 30013, Taiwan. Electronic address: lswwg@life.nthu.edu.tw.
authors.forename: Wen-Guey
authors.initials: WG
authors.lastname: Wu
mesh_terms: D051722:ADAM Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D020816:Amino Acid Motifs; D000818:Animals; D000077214:Becaplermin; D002448:Cell Adhesion; D002465:Cell Movement; D004546:Elapid Venoms / Q000201:enzymology*; D017815:Elapidae; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D039302:Integrin alphaVbeta3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008024:Ligands; D051379:Mice; D062105:Molecular Docking Simulation; D041681:NIH 3T3 Cells; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D020574:Proto-Oncogene Proteins c-sis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D030162:Reptilian Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012995:Solubility; D020349:Surface Plasmon Resonance; D013624:Taiwan
chemical_list: D004546:Elapid Venoms; D055767:Immobilized Proteins; D039302:Integrin alphaVbeta3; D008024:Ligands; D010446:Peptide Fragments; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; D030162:Reptilian Proteins; D000077214:Becaplermin; D051722:ADAM Proteins
keywords: Atragin; Cell migration; Integrin; Kaouthiagin-like; Snake venom metalloproteinases (SVMPs); Taiwan cobra venom