title: Electrophysiological Characterization of the Antarease Metalloprotease from Tityus serrulatus Venom.
abstract: Scorpions are among the oldest venomous living organisms and the familyButhidaeis the largest and most medically relevant one. Scorpion venoms include many toxic peptides, but recently, a metalloprotease fromTityus serrulatuscalled antarease was reported to be capable of cleaving VAMP2, a protein involved in the neuroparalytic syndromes of tetanus and botulism. We have produced antarease and an inactive metalloprotease mutant in a recombinant form and analyzed their enzymatic activity on recombinant VAMP2 in vitro and on mammalian and insect neuromuscular junction. The purified recombinant antarease paralyzed the neuromuscular junctions of mice and ofDrosophila melanogasterwhilst the mutant was inactive. We were unable to demonstrate any cleavage of VAMP2 under conditions which leads to VAMP proteolysis by botulinum neurotoxin type B. Antarease caused a reduced release probability, mainly due to defects upstream of the synaptic vesicles fusion process. Paired pulse experiments indicate that antarease might proteolytically inactivate a voltage-gated calcium channel.
authors.affiliation: Dipartimento di Scienze Biomediche and Istituto CNR di Neuroscienze, Università di Padova, Via Ugo Bassi 58/B, 35121 Padova, Italy. irenezornetta@gmail.com.
authors.forename: Irene
authors.initials: I
authors.lastname: Zornetta
authors.affiliation: Dipartimento di Scienze Biomediche and Istituto CNR di Neuroscienze, Università di Padova, Via Ugo Bassi 58/B, 35121 Padova, Italy. scorzetomichele@gmail.com.
authors.forename: Michele
authors.initials: M
authors.lastname: Scorzeto
authors.affiliation: Laboratório de Venenos e Toxinas Animais, Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, MG, Brazil. reispvm@gmail.com.
authors.forename: Pablo Victor
authors.initials: PV
authors.lastname: Mendes Dos Reis
authors.affiliation: Laboratório de Venenos e Toxinas Animais, Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, MG, Brazil. lima.mariaelena@gmail.com.
authors.forename: Maria E
authors.initials: ME
authors.lastname: De Lima
authors.affiliation: Dipartimento di Scienze Biomediche and Istituto CNR di Neuroscienze, Università di Padova, Via Ugo Bassi 58/B, 35121 Padova, Italy. cesare.montecucco@gmail.com.
authors.forename: Cesare
authors.initials: C
authors.lastname: Montecucco
authors.affiliation: Dipartimento di Scienze Biomediche and Istituto CNR di Neuroscienze, Università di Padova, Via Ugo Bassi 58/B, 35121 Padova, Italy. aram.megighian@unipd.it.
authors.forename: Aram
authors.initials: A
authors.lastname: Megighian
authors.affiliation: Dipartimento di Scienze Biomediche and Istituto CNR di Neuroscienze, Università di Padova, Via Ugo Bassi 58/B, 35121 Padova, Italy. ornella.rossetto@unipd.it.
authors.forename: Ornella
authors.initials: O
authors.lastname: Rossetto
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000633:toxicity*; D003964:Diaphragm; D004331:Drosophila melanogaster / Q000187:drug effects / Q000502:physiology; D005071:Evoked Potentials / Q000187:drug effects; D007814:Larva; D008297:Male; D045726:Metalloproteases / Q000633:toxicity*; D051379:Mice; D009119:Muscle Contraction / Q000187:drug effects; D009466:Neuromuscular Blocking Agents / Q000633:toxicity*; D009469:Neuromuscular Junction / Q000187:drug effects / Q000502:physiology; D010791:Phrenic Nerve / Q000187:drug effects / Q000502:physiology; D011994:Recombinant Proteins / Q000633:toxicity; D050600:SNARE Proteins / Q000378:metabolism; D012604:Scorpion Venoms; D012605:Scorpions
chemical_list: D060829:Arthropod Proteins; D009466:Neuromuscular Blocking Agents; D011994:Recombinant Proteins; D050600:SNARE Proteins; D012604:Scorpion Venoms; D045726:Metalloproteases
keywords: Zn-metalloprotease; antarease; botulinum neurotoxins; scorpion; vesicle associated membrane protein 2 (VAMP2)