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Supplemental Information Membrane-proximal tryptophans of synaptobrevinII stabilize priming of secretory vesicles 1 1 1 1 Maria Borisovska , Yvonne N. Schwarz , Madhurima Dhara , Antonio Yarzagaray , Sandra 1 2 2 1 1 Hugo , Daniele Narzi , Shirley W.I. Siu , Jaideep Kesavan , Ralf Mohrmann , Rainer A. 2 1 Böckmann and **Dieter Bruns 1 Institute for Physiology School of Medicine University of Saarland, Germany 2 Computational Biology Department of Biology University of Erlangen-Nürnberg, Germany Supplementary Figure 1 Figure S1. Substitution or deletion of the membrane-proximal Trp-residues of sybII leaves the spike phase of transmitter discharge and the pre-spike signal (‘foot’) unchanged. Properties of amperometric spikes preceded by a ‘foot’ signal probably arise from events close to the detector as indicated by their fast kinetics. Data were collected from sybII (n=24), -WW (n=18), WW/AA (n=15) and WW/SS (n=15) expressing dko cells. Note that neither spike properties nor pre-spike-properties are affected by mutating the TRP-residues. The start of the ‘foot’ signal is defined as the time point where the current amplitude exceeds two times the standard deviation of the average baseline noise and it ends at the inflection point (determined from maximum of the current second derivative) between the slowly increasing ‘foot’ signal and the rapidly increasing spike current. For ‘foot’ flicker analysis, the current derivative was again filtered at 1.2 kHz and fluctuations exceeding the threshold of ± 6 pA/ms (~4 times SD of baseline noise) were counted. The number of suprathreshold fluctuations divided by the corresponding ‘foot’ duration defines the fluctuation frequency. Values for spike, pre-spike amplitude and percentage of pre-spike events are given as mean (± s.e.m.). Other values are given as average median determined from the parameter’s frequency distribution for each cell.