caa a22 4500 463254104 CHVBK 20180405153349.0 cr unu---uuuuu 170326e20070101xx s 000 0 eng 10.1007/s10571-006-9123-z doi (NATIONALLICENCE)springer-10.1007/s10571-006-9123-z Leftward Shift in the Voltage-Dependence for Ca2+ Currents Activation Induced by a New Toxin from Phoneutria reidyi (Aranae, Ctenidae) Venom [Elektronische Daten] [L. Vieira, A. Pimenta, M. Richardson, M. Bemquerer, H. Reis, J. Cruz, M. Gomez, M. Santoro, R. Ferreira-de-Oliveira, S. Figueiredo, T. Snutch, M. Cordeiro] Summary: Various neurotoxins have been described from the venom of the Brazilian spider Phoneutria nigriventer, but little is known about the venoms of the other species of this genus. In the present work, we describe the purification and some structural and pharmacological features of a new toxin (PRTx3-7) from Phoneutria reidyi that causes flaccid paralysis in mice. The observed molecular mass (4627.26Da) was in accordance with the calculated mass for the amidated form of the amino acid sequence (4627.08Da). The presence of an α-amidated C-terminus was confirmed by MS/MS analysis of the C-terminal peptide, isolated after enzymatic digestion of the native protein with Glu-C endoproteinase. The purified protein was injected (intracerebro-ventricular) into mice at dose levels of 5μg/mouse causing immediate agitation and clockwise gyration, followed by the gradual development of general flaccid paralysis. PRTx3-7 at 1μM inhibited by 20% the KCl-induced increase on [Ca2+]i in rat brain synaptosomes. The HEK cells permanently expressing L, N, P/Q and R HVA Ca2+ channels were also used to better characterize the pharmacological features of PRTx3-7. To our surprise, PRTx3-7 shifted the voltage-dependence for activation towards hyperpolarized membrane potentials for L (−4mV), P/Q (−8mV) and R (−5mV) type Ca2+ currents. In addition, the new toxin also affected the steady state of inactivation of L-, N- and P/Q-type Ca2+ currents. Springer Science+Business Media, LLC, 2006 Phoneutria reidyi nationallicence spider venom nationallicence neurotoxin nationallicence amidated C-terminus nationallicence Ca2+-channel toxin nationallicence Vieira L. Laboratório de Neurofarmacologia, Departamento de Farmacologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut Pimenta A. Laboratório de Venenos e Toxinas Animais, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, Campus Pampulha, Caixa Postal 486, CEP 31270-901, Belo Horizonte, MG, Brazil aut Richardson M. Centro de Pesquisa Professor Carlos Ribeiro Diniz, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil aut Bemquerer M. Laboratório de Venenos e Toxinas Animais, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, Campus Pampulha, Caixa Postal 486, CEP 31270-901, Belo Horizonte, MG, Brazil aut Reis H. Laboratório de Neurofarmacologia, Departamento de Farmacologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut Cruz J. Laboratório de Membranas Excitáveis, Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut Gomez M. Laboratório de Neurofarmacologia, Departamento de Farmacologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut Santoro M. Núcleo de Biomoléculas, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut Ferreira-de-Oliveira R. Centro de Pesquisa Professor Carlos Ribeiro Diniz, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil aut Figueiredo S. Departamento de Ciências Biológicas, Universidade Federal de Espírito Santo, Vitória, ES, Brazil aut Snutch T. Biotechnology Laboratory, University of British Columbia, Vancouver, BC, Canada aut Cordeiro M. Centro de Pesquisa Professor Carlos Ribeiro Diniz, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil aut Cellular and Molecular Neurobiology Springer US; http://www.springer-ny.com 27/1(2007-01-01), 129-146 0272-4340 27:1<129 2007 27 10571 https://doi.org/10.1007/s10571-006-9123-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10571-006-9123-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Vieira L. Laboratório de Neurofarmacologia, Departamento de Farmacologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 700 1- Pimenta A. Laboratório de Venenos e Toxinas Animais, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, Campus Pampulha, Caixa Postal 486, CEP 31270-901, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 700 1- Richardson M. Centro de Pesquisa Professor Carlos Ribeiro Diniz, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 700 1- Bemquerer M. Laboratório de Venenos e Toxinas Animais, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, Campus Pampulha, Caixa Postal 486, CEP 31270-901, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 700 1- Reis H. Laboratório de Neurofarmacologia, Departamento de Farmacologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 700 1- Cruz J. Laboratório de Membranas Excitáveis, Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 700 1- Gomez M. Laboratório de Neurofarmacologia, Departamento de Farmacologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 700 1- Santoro M. Núcleo de Biomoléculas, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 700 1- Ferreira-de-Oliveira R. Centro de Pesquisa Professor Carlos Ribeiro Diniz, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 700 1- Figueiredo S. Departamento de Ciências Biológicas, Universidade Federal de Espírito Santo, Vitória, ES, Brazil aut NATIONALLICENCE 700 1- Snutch T. Biotechnology Laboratory, University of British Columbia, Vancouver, BC, Canada aut NATIONALLICENCE 700 1- Cordeiro M. Centro de Pesquisa Professor Carlos Ribeiro Diniz, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil aut NATIONALLICENCE 773 0- Cellular and Molecular Neurobiology Springer US; http://www.springer-ny.com 27/1(2007-01-01), 129-146 0272-4340 27:1<129 2007 27 10571 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer