Cesare Montecucco, Aram Megighian, and Ornella Rossetto. Figure S1. Paired pulse facilitation. Example of paired pulse facilitation following paired stimulation.
Toxins 2017, 9, 81; doi:10.3390/toxins9030081
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Supplementary Materials: Electrophysiological Characterization of the Antarease Metalloprotease from Tityus serrulatus Venom Irene Zornetta, Michele Scorzeto, Pablo Victor Mendes Dos Reis, Maria E. De Lima, Cesare Montecucco, Aram Megighian, and Ornella Rossetto
Figure S1. Paired pulse facilitation. Example of paired pulse facilitation following paired stimulation (black arrows) of segmental nerve innervating fibre 6 of abdominal segment 3 in third instar larva body wall preparation. The second Excitatory postsynaptic potential (EPP) is augmented with respect to the first EPP. Interstimulus interval is 75 ms (gray double arrow). Calibration bar on the right side of the figure. 1
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165 125 93 72 57 42 31 24 18 15
Figure S2. Purification of recombinant antarease. T. serrulatus E162A antarease was produced as recombinant protein in E. coli and subjected to SDS‐PAGE. Lane 1: molecular weight markers; lane 2: total protein extract; lanes 3 and 4: washes; lanes 5: surnatant after sarcoside 0.5%; lanes 6‐9: eluted fractions from the affinity chromatography containing E162A antarease (arrow).
Toxins 2017, 9, 81; doi:10.3390/toxins9030081
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Relative amplitude
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Vehicle AntaeraseE162A mutant
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Ve hi cl e
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as eE 16 2A
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as e
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Figure S3. EJP Amplitude 500 s after addition of recombinant proteins. Mean ± Standard Deviation of EJP Amplitudes of 4 experiments for each experimental condition, measured 500 s after addition of recombinant proteins or vehicle to the bath. EJP amplitudes are relative to the mean of the last 10 EJPs recorded, before adding recombinant proteins or vehicle to the bath, in the same third instar D. melanogaster larval muscle 6/7 of A3/A4 segment. Microlectrode remained intracellularly placed throughout the entire experiment.
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A 50 ntae An 50 tae 1 An ras A r 00 ta e 10 nta ase A era s E n 0 An era 16 tae e r s 2 ta eE A as er e as 162 mu eE A tan 16 mu t 2 A ta m nt 20 uta V nt 50 ehic V le 10 eh 0 icl Ve e hi cl e
0.0
Figure S4. P2/P1 ratio of double pulse stimulation. Mean ± Standard Deviation of Pulse 2/ Pulse 1 (P2/P1) of 4 experiments for each experimental condition, measured 650 s after addition of recombinant proteins or vehicle to the bath, when Antaerase was fully effective.
