DOI: 10.5061/dryad.0zpc866vt
title: NMR data in 'Ca2+-dependent release of Synaptotagmin-1 from the SNARE complex on phosphatidylinositol 4,5-bisphosphate-containing membranes' by Voleti, Jaczynska and Rizo, eLife 2020

description: The Ca2+ sensor synaptotagmin-1 and the SNARE complex cooperate to trigger
 neurotransmitter release. Structural studies elucidated three distinct
 synaptotagmin-1-SNARE complex binding modes involving ‘polybasic’,
 ‘primary’ and ‘tripartite’ interfaces of synaptotagmin-1. We investigated
 these interactions using NMR and fluorescence spectroscopy.
 Synaptotagmin-1 binds to the SNARE complex through the polybasic and
 primary interfaces in solution. Ca2+-free synaptotagmin-1 binds to SNARE
 complexes anchored on PIP2-containing nanodiscs. R398Q/R399Q and
 E295A/Y338W mutations at the primary interface, which strongly impair
 neurotransmitter release, disrupt and enhance synaptotagmin-1-SNARE
 complex binding, respectively. Ca2+ induces tight binding of
 synaptotagmin-1 to PIP2-containing nanodiscs, releasing
 synaptotagmin-1-SNARE interactions. Specific effects of mutations in the
 polybasic region on Ca2+-dependent synaptotagmin-1-PIP2-membrane
 interactions correlate with their effects on release. Our data suggest
 that synaptotagmin-1 binds to the SNARE complex through the primary
 interface and that Ca2+ releases this interaction, inducing PIP2/membrane
 binding and allowing cooperation between synaptotagmin-1 and the SNAREs in
 membrane fusion to trigger release.
description: The dataset consists of <sup>1</sup>H
 <sup>15</sup>N TROSY HSQC and
 <sup>1</sup>H<sup>13</sup> HMQC spectra acquired
 to investigate interactions of Synaptotagmin-1 with
 Complexin-1(26-83)-SNARE complex in solution. All NMR
 spectra in the data set were acquired at 25°C on Agilent DD2 spectrometers
 operating at 600 or 800 MHz and equipped with cold probes. Detailed sample
 preparation is described in the methods section of Voleti et al, 2020. NMR
 data were processed with NMRPipe (Delaglio et al., 1995) and analyzed with
 NMRView (Johnson and Blevins, 1994). The data sets are
 organized in main directories that correspond to the figures of Voleti et
 al, 2020, i.e. all the files that were used for generating Figure 1 are
 included in main directory “Fig1”. Within each of these directories there
 is one directory for each titration included in the corresponding figure
 and within each titration directory there is one directory for each
 spectrum acquired during the titration. These directories have extension
 .fid and their name specifies the experimental conditions (including
 mutations and protein concentration). Each of these directories includes
 files that contain the raw NMR data (fid), the acquisition parameters
 (procpar), a log file (log), a text file with a more detailed description
 of experimental conditions (text), macros to convert from Agilent format
 to NMRPipe format (fid.com) and to process the data with NMRPipe
 (jrtrgood2c.com), an intermediate processing file (tr.ft), the processed
 NMR spectrum in NMRPipe format (tr.ft2) and the same spectrum converted to
 NMR view format (same name as the directory but with extension .nv). For
 each titration, there is also a directory called nvs that contains all the
 .nv files. To increase the signal to noise ratio, some of the experiments
 were acquired multiple times and added. For added experiments, an
 additional directory was created (directory name includes _add) and those
 nv files were used for further analysis. The spectra included in the
 addition are indicated by the numbers at the end of the filename. Below is
 a summary of the main directories and the directories corresponding to
 each titration. NMR figures list:
 Fig1
 TRHS_201803_SL-IM_C2B_Syt1_WT_mcc-cpx26-83_titration_EDTA   Fig1-S2 TRHS_201803_SL-IM_C2B_Syt1_WT_mcc-cpx26-83_titration_CaCl2   Fig2 TRHS_201805_SL-IM_C2B_Syt1_RRQQ_mcc-cpx26-83_titration_EDTA TRHS_20190505_SL-IM_C2B_Syt1_RKEE_mcc-cpx26-83_titration_EDTA TRHS_201907_SL-IM_C2B_Syt1_RKRREEQQ_mcc-cpx26-83_titration_EDTA   Fig2-S1 TRHS_201806_SL-IM_C2B_Syt1_R399Q_mcc-cpx26-83_titration_EDTA TRHS_201806_SL-IM_C2B_Syt1_R398Q_mcc-cpx26-83_titration_EDTA   Fig3 TRHS_201805_2H15N_Cpx2683mcc_S1C2B_WT_titration_EDTA TRHS_201904_2H15N_Cpx2683mcc_S1C2B_RKRREEQQ_titration_EDTA   Fig3-S1 201805_SL-IM_C2B_Syt1_LLQQ_mcc-cpx2683_titration_EDTA   Fig4 TRHS_201906_SL-IM_C2B_Syt1_EYAW_mcc-cpx26-83_titration_EDTA   Fig4-S1 HMQC_201803_SL-IM_C2B_Syt1_WT_mcc-cpx26-83_titration_EDTA HMQC_201905_SL-IM_C2B_Syt1_RKEE_mcc-cpx26-83_titration_EDTA HMQC_201906_SL-IM_C2B_Syt1_EYAW_mcc-cpx26-83_titration_EDTA HMQC_201805_SL-IM_C2B_Syt1_RRQQ_mcc-cpx26-83_titration_EDTA HMQC_201907_SL-IM_C2B_Syt1_RKRREEQQ_mcc-cpx26-83_titration_EDTA
description: To view the files, please download the file
 Voleti_Jaczynska_NMR_data.tar.gz and use the following commands:
 gunzip
 Voleti_Jaczynska_NMR_data.tar.gz tar xvf
 Voleti_Jaczynska_NMR_data.tar Processed files with
 extension .nv or .ft2 are ready to be viewed in NMRViewJ or NMRDraw,
 respectively. In case of any questions, please do not
 hesitate to contact the authors.