title: Structural analysis of trimeric phospholipase A2 neurotoxin from the Australian taipan snake venom.
abstract: Snake pre-synaptic neurotoxins endowed with phospholipase A(2) activity are potent inducers of paralysis through the specific disruption of the neuromuscular junction pre-synaptic membrane and represent a valuable tool for investigating neuronal degeneration and recovery. They have different structural complexity and a wide range of lethal potency and enzymatic activity, although they share a similar mechanism of action. Although no correlation has been reported between neurotoxicity and enzymatic activity, toxicity increases with structural complexity and phospholipase A(2) oligomers show 10-fold lower LD(50) values compared to their monomeric counterparts. To date, no structural study has been performed on multimeric SPANs with the aim of shedding light on the correlation between structural complexity and neurotoxicity. In the present study, we investigated the structure of taipoxin, a trimeric phospholipase A(2) neurotoxin, as well as that of its subunits, by X-ray crystallography and small angle X-ray scattering analysis. We present the high-resolution structure of two isoforms of the taipoxin β subunit, which show no neurotoxic activity but enhance the activity of the other subunits in the complex. One isoform shows no structural change that could justify the lack of activity. The other displays three point mutations in critical positions for the catalytic activity. Moreover, we designed a model for the quaternary structure of taipoxin under physiological conditions, in which the three subunits are organized into a flat holotoxin with the substrate binding sockets exposed on the same side of the complex, which suggests a role for this interface in the toxin-membrane interaction.
authors: Department of Biological Chemistry, University of Padova, Italy.
authors: Laura
authors: L
authors: Cendron
authors: Ivan
authors: I
authors: Mičetić
authors: Patrizia
authors: P
authors: Polverino de Laureto
authors: Marco
authors: M
authors: Paoli
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D018360:Crystallography, X-Ray; D004546:Elapid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000633:toxicity; D017815:Elapidae; D007527:Isoenzymes / Q000737:chemistry / Q000633:toxicity; D007928:Lethal Dose 50; D008958:Models, Molecular; D008969:Molecular Sequence Data; D054467:Phospholipases A2 / Q000737:chemistry* / Q000633:toxicity; D011487:Protein Conformation; D053838:Scattering, Small Angle; D017386:Sequence Homology, Amino Acid
chemical_list: D004546:Elapid Venoms; D007527:Isoenzymes; C015523:taipoxin; D054467:Phospholipases A2