Data from: Synaptotagmin-1 serves as a primary Zn2+ sensor to mediate spontaneous neurotransmitter release under pathological conditions
Data files
Jan 30, 2026 version files 22.59 MB
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README.md
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Source_data.zip
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Abstract
This dataset provides the source data for the paper titled "Synaptotagmin-1 serves as a primary Zn²⁺ sensor to mediate spontaneous neurotransmitter release under pathological conditions." Data includes electrophysiological patch-clamp recordings of miniature excitatory postsynaptic currents (mEPSCs), spontaneous EPSCs (sEPSCs), and evoked EPSCs, along with single-vesicle and ensemble lipid-mixing assays (in vitro reconstitution system) measuring fusion probability (%), cumulative probability, and lipid-mixing efficiency (FRET). The dataset also covers zinc-protein or membrane-binding assays evaluating binding affinity and membrane-binding capacity, as well as vesicle docking imaging via TIRF microscopy, which quantifies docking-related fluorescence intensity and the number of docked vesicles. Data collection and analysis were conducted collaboratively across multiple institutions. Electrophysiological patch-clamp recordings and vesicle docking imaging were performed at the Key Laboratory of Xi'an Jiaotong University, while the remaining data were collected at the relevant experimental facilities of Sichuan University.
Dataset DOI: 10.5061/dryad.np5hqc05k
Description of the data and file structure
Files and variables
Description of the data and file structure
This dataset (Source_data.zip) comprises four integrated experimental modules: electrophysiology, in vitro vesicle fusion reconstitution, zinc ion binding and membrane interaction analysis, and total internal reflection fluorescence (TIRF) microscopy imaging. Electrophysiological recordings from hippocampal neurons and slices—including miniature excitatory postsynaptic currents (mEPSC), spontaneous EPSC (sEPSC), and evoked EPSC—as well as vesicle docking imaging via TIRF microscopy, were conducted at the Key Laboratory of Xi'an Jiaotong University. In vitro single-vesicle and ensemble lipid-mixing assays, along with zinc-protein binding and membrane co-sedimentation experiments, were performed at relevant experimental facilities of Sichuan University. All raw data and statistical results are systematically organized by figure number in corresponding worksheets within a single Excel file (Source data.xlsx), ensuring full correspondence and traceability between data and figures. Analysis was carried out using GraphPad Prism 8.0, ImageJ, and OriginPro 2024. Detailed experimental procedures and analytical workflows are provided in the "Methods" section and supplementary materials of the associated paper.
Code/software
File: Source data.xlsx
Figure1c, d, f, h and i
Description: This file provides electrophysiology data from hippocampal neurons measuring spontaneous(mEPSC) and evoked (EPSC) release frequencies and amplitudes under varying Zn²⁺ concentrations.
Variables
Figure 1c and d :
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- mEPSC frequency (Hz) – Frequency of miniature excitatory postsynaptic currents in wild-type Syt1 neurons (unit: hertz).
- mEPSC amplitude (pA) – Amplitude of miniature excitatory postsynaptic currents in wild-type Syt1 neurons (unit: picoampere).
Figure 1f :
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- Evoked EPSC amplitude (pA) – Amplitude of evoked excitatory postsynaptic currents in wild-type Syt1 neurons (unit: picoampere).
Figure 1h and i :
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- mEPSC frequency (Hz) – Frequency of miniature excitatory postsynaptic currents in Syt1 conditional knockout neurons (unit: hertz).
- mEPSC amplitude (pA) – Amplitude of miniature excitatory postsynaptic currents in Syt1 conditional knockout neurons (unit: picoampere).
Figure 2b,c, d, e, f and g
Description: This file provides the probability distribution and probability of vesicle fusion mediated by different conditions.
Variables
Figure 2b and c :
- Time (s) – Recording time point (unit: seconds).
- 500 μM Ion-dependent fusion probability – probability distribution of fusion events per second, recorded over 1 minute in the presence of ions(Sr²⁺、Mg²⁺、Ca²⁺、Zn²⁺) (unitless, Probability scale: Normalized cumulative probability (0-1 )).
- Cumulative probability – Cumulative probability of ions(Sr²⁺、Mg²⁺、Ca²⁺、Zn²⁺)-dependent vesicle fusion events (unitless).
- Zn²⁺ – Reaction progress treatmented by Zinc ions.
- Ca²⁺ – Reaction progress treatmented by Calcium ions .
- Mg²⁺ – Reaction progress treatmented by Magnesium ions.
- Sr²⁺ – Reaction progress treatmented by Strontium ions.
Figure 2d and e :
- Time (s) – Recording time point (unit: seconds).
- Zn²⁺ concentrations (µM)– 0、10、20、50、100 and 500 µM zinc ion (unit: micromolar).
- Zn²⁺ -dependent fusion probability – probability distribution of fusion events per second, recorded over 1 minute in the presence of ions (unitless, Probability scale: Normalized cumulative probability (0-1 )).
- Cumulative probability – Cumulative probability of different Zn²⁺ concentrations(0、10、20、50、100 and 500 µM)-dependent vesicle fusion events (unitless).
Figure 2f and g :
- Time (s) – Recording time point (unit: seconds).
- 500 µM Zn²⁺-dependent fusion probability – probability distribution of fusion events per second, recorded over 1 minute in the presence of Syt1 and its mutants(Syt1_WT、Syt1_C2AB、Syt1_4A、Syt1_4W) (unitless, Probability scale: Normalized cumulative probability (0-1 )).
- Syt1_WT – wild-type synaptotagmin-1 reconstituted into vesicles reconstituted into vesicles.
- Syt1_C2AB -C2 domain calcium-binding mutant reconstituted into vesicles reconstituted into vesicles.
- Syt1_4A - Syt1 membrane-insertion mutant reconstituted into vesicles reconstituted into vesicles.
- Syt1_4W - Syt1 membrane-insertion mutant reconstituted into vesicles reconstituted into vesicles.
- Cumulative probability – 500 µM Zn²⁺-dependent fusion probability of Syt1 and its mutants(Syt1_WT、Syt1_C2AB、Syt1_4A、Syt1_4W) (unitless).
Figure3b,c, f and g
Description: This file provides fluorescence spectroscopy data from FluoZin-1 assays for Zn²⁺ binding to the Syt1 C2AB and variants.
Variables
Figure 3b, c and f :
- None – Buffer-only control (no protein).
- C2AB – Full cytoplasmic domain fragment of synaptotagmin-1.
- C2AB(Zn²⁺) – C2AB fragment pre-incubated with zinc ions.
- C2A – Isolated C2A domain fragment of synaptotagmin-1.
- C2B – Isolated C2B domain fragment of synaptotagmin-1.
- C2AB – C2AB mutant with disrupted calcium-binding sites.
- C2AB_3M – C2AB mutant with disrupted zinc-binding residues (D261A/E386A/H389A).
- Em. Wavelength (nm) – Emission peak wavelength in fluorescence measurements (unit: nanometers).
- Intensity (Counts/s/µA) – Fluorescence signal intensity measured at the corresponding peak wavelength (unit: arbitrary units(normalized fluorescence intensity)).
- SE – Standard Error of the Mean for Intensity(unit: Same as the Intensity (unitless)).
Figure 3g:
- Concentration (µM) – Protein concentration in titration experiments ( unit: micromolar).
- Binding Affinity (Kd) – Dissociation constant from titration curves ( unit: nanomolar or micromolar).
- C2AB: Full cytoplasmic domain fragment of synaptotagmin-1.
- C2AB_3M – C2AB mutant with disrupted zinc-binding residues (D261A/E386A/H389A).
- Intensity (a.u.) – Fluorescence signal intensity measured at the corresponding peak wavelength (unit: arbitrary units(normalized fluorescence intensity)).
- SE – Standard Error of the Mean for Intensity(unit: Same as the Intensity (unitless)).
Figure 4a, b, d, e, g and h
Description: This file provides probability of vesicle fusion mediated by Syt1_WT or Syt1_3M and electrophysiology data from Syt1 cKO hippocampal neurons rescued by Syt1_WT or Syt1_3M measuring spontaneous(mEPSC).
Variables
Figure 4a and b:
- Time (s) – Recording time point (unit: seconds).
- 500 nM or 500 µM Zn²⁺-dependent fusion probability – probability distribution of fusion events per second, recorded over 1 minuten the presence of Syt1_WT or Syt1_3M (unitless, Probability scale: Normalized cumulative probability (0-1 )).
- Syt1_WT – wild-type Syt1 protein reconstituted into vesicles reconstituted into vesicles.
- Syt1_3M – Syt1 zinc-binding mutant protein reconstituted into vesicles reconstituted into vesicles.
- Cumulative probability – Cumulative probability of 500 nM or 500 µM Zn²⁺-dependent vesicle fusion events (unitless)
Figure 4d, e, g and h
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- cKO + Syt1_WT – Syt1 cKO hippocampal neurons rescued by Syt1_WT.
- cKO + Syt1_3M –Syt1 cKO hippocampal neurons rescued by Syt1_3M.
- mEPSC frequency (Hz) – Frequency of miniature excitatory postsynaptic currents in wild-type Syt1 neurons (unit: hertz).
- mEPSC amplitude (pA) – Amplitude of miniature excitatory postsynaptic currents in wild type Syt1 neurons (unit: picoampere).
Figure 5d, e and f
Description: This file provides gel images and precipitation fluorescence spectra related to the study of the binding ability of Zn²⁺ to membranes containing different compositions of phosphatidylserine (PS) and phosphatidylinositol (PIP2).
Variables
- (0, 5, 10 and 15) PS concentration (%) – Phosphatidylserine content in lipid vesicles (unit: percentage).
- 1 PIP₂ concentration (%) – Phosphatidylinositol 4,5-bisphosphate content in lipid vesicles (unit: percentage).
- 15 PS + 1 PIP₂ concentration (%) – Phosphatidylserine and Phosphatidylinositol 4,5-bisphosphate content in lipid vesicles (unit: percentage).
- Zn²⁺ concentration (µM) – Concentration of zinc ions in the experimental solution (unit: micromolar).
- C2AB – Cytosolic tandem C2-domains of synaptotagmin-1.
- C2AB_3M – Zinc-binding deficient triple mutant (D261A/E386A/H389A).
- S – Supernatant in fraction.
- P – Pellet in fraction.
- Intensity (Counts/s/µA) – Fluorescence signal intensity measured at the corresponding peak wavelength (unit: arbitrary units(normalized fluorescence intensity)).
- Normalized P/S – Normalized ratio of protein bound to the pellet fraction versus remaining in the supernatant fraction.
- KDa– Kilo-Dalton, measuring protein mass.
Figure 6c, d and f
Description: This file provides on the quantity of vesicle docking promoted by Zn²⁺ in vitro and the changes in cellular imaging fluorescence intensity.
Variables
Figure 6c and d:
- WT – wild-type hippocampal neurons.
- Syt1cKO – Syt1 cKO hippocampal neurons.
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- Time (s) – Elapsed time from the start of the recording.(unit: seconds (s)).
- Intensity – The fluorescence intensity trace for NH₄Cl perfusion pulse.
- I₀ – The fluorescence intensity trace for the first NH₄Cl perfusion pulse (before Zn²+ application). The peak value in this phase is defined as I₀(unit: Arbitrary Fluorescence Units (a.u.)).
- IZn²+ – The fluorescence intensity trace from neurons treated with 200 µM Zn²⁺. The trace shows the response to NH₄Cl after zinc application. The peak in this phase is defined as I_Zn²⁺(unit: Arbitrary Fluorescence Units (a.u.)).
- Intensity ratio (I₀ /IZn²+ ) – dividing the maximal fluorescence intensity obtained during the first ammonium chloride (NH₄Cl) perfusion (I₀) by the maximal fluorescence intensity obtained during the second NH₄Cl perfusion performed after the application of Zn²⁺ (I_Zn²⁺).
Figure 6f:
- Syt1_WT – Wild-type full-length Syt1 reconstituted into vesicles.
- Syt1_QM –synaptotagmin-1 primary interface with the SNARE complex region mutant reconstituted into vesicles.
- Syt1_QQQ – synaptotagmin-1 polybasic region mutant mutant reconstituted into vesicles.
- Syt1_3M –Syt1 zinc-binding mutant reconstituted into vesicles reconstituted into vesicles.
- Zn²⁺ concentration (µM) – Treatment with 0 or 500 µM zinc ion concentrations (unit: micromolar).
- Counts – the number of SV vesicles bound to PM vesicles.
Supplementary Data 1b and c
Description: This file provides electrophysiology data from hippocampal neurons measuring spontaneous(sEPSC) release frequencies and amplitudes under varying Zn²⁺ concentrations.
Variables
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- sEPSC frequency (Hz) – Frequency of spontaneous excitatory postsynaptic Current in wild-type Syt1 neurons (unit: hertz).
- sEPSC amplitude (pA) – Amplitude of spontaneous excitatory postsynaptic currents in wild-type Syt1 neurons (unit: picoampere).
Supplementary Data 2b and c
Description: This file provides electrophysiology data from hippocampal slices measuring spontaneous(mEPSC) release frequencies and amplitudes under varying Zn²⁺ concentrations.
Variables
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- mEPSC frequency (Hz) – Frequency of miniature excitatory postsynaptic currents in hippocampal slices (unit: hertz).
- mEPSC amplitude(pA) – Amplitude of miniature excitatory postsynaptic currents in hippocampal slices (unit: picoampere).
Supplementary Data 3b, c and e
Description: This file provides electrophysiology data from hippocampal neurons measuring spontaneous(mEPSC) and evoked (EPSC) release frequencies and amplitudes in the absence of Ca2+ .
Variables
Supplementary Data 3b and c :
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- Ca²⁺ concentration (µM) – Treatment with different extracellular calcium ion concentrations (unit: micromolar).
- mEPSC frequency (Hz) – Frequency of miniature excitatory postsynaptic currents in wild-type Syt1 neurons (unit: hertz).
- mEPSC amplitude (pA) – Amplitude of miniature excitatory postsynaptic currents in wild-type Syt1 neurons (unit: picoampere).
Supplementary Data 3e :
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- Ca²⁺ concentration (µM) – Treatment with different extracellular calcium ion concentrations (unit: micromolar).
- Evoked EPSC amplitude (pA) – Amplitude of evoked excitatory postsynaptic currents in wild-type Syt1 neurons (unit: picoampere).
Supplementary Data 4b and c
Description: This file provides electrophysiology data from hippocampal slices measuring spontaneous(sEPSC) release frequencies and amplitudes under varying Zn²⁺ concentrations.
Variables
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- mEPSC frequency (Hz) – Frequency of spontaneous spontaneous postsynaptic currents in Syt1 conditional knockout neurons (unit: hertz).
- mEPSC amplitude (pA) – Amplitude of spontaneous spontaneous postsynaptic currents in Syt1 conditional knockout neurons (unit: picoampere).
Supplementary Data 5a and b
Description: This file provides the distribution and efficiency changes of Zn²⁺-dependent vesicle fusion with varying protein types, using single-vesicle fusion assay.
Variables
- Time (s) – Recording time point (unit: seconds).
- Zn²⁺-dependent fusion probability – probability distribution of fusion events per second, recorded over 1 minute in the presence of ions (unitless, Probability scale: Normalized cumulative probability (0-1 )).
- Cumulative probability – Cumulative probability of Zn²⁺-dependent vesicle fusion events by different protein types (unitless).
- Syt1_WT – Wild-type Synaptotagmin-1 reconstituted into vesicles. This is the positive control with all necessary components.
- C2AB – Only the cytosolic tandem C2 domains of Syt1.
- No Syt1 – Vesicles reconstituted without Synaptotagmin-1.
- No VAMP2 – Vesicles lacking Vesicle-Associated Membrane Protein 2 (Synaptobrevin).
- No SNAP-25 – Vesicles lacking Synaptosomal-Associated Protein 25 kDa.
Supplementary Data 6b and c
Description: This file provides the maximum fusion probability(FRET-based) and efficiency changes of vesicle fusion dependent on different divalent ion,using lipid mixing assay
Variables
- Time (s) – Recording time point (unit: seconds)
- Zn²⁺ – Reaction progress treated by Zinc ions
- Ca²⁺ – Reaction progress treated by Calcium ions
- Mg²⁺ – Reaction progress treated by Magnesium ions
- Sr²⁺ – Reaction progress treated by Strontium ions
- Maximum fusion probability – Measured at the corresponding ions(unit: Unitless probability (range 0 to 1, where 0 = no fusion, 1 = all vesicles fuse).)
- SE – Standard Error of the Mean for the Maximum Fusion Probability(unit: Same as the probability (unitless))
Supplementary Data 7a and b
Description: This file provides the maximum fusion probability(FRET-based) and efficiency changes of vesicle fusion dependent on Zn2+ and Ca2+,using lipid mixing assay.
Variables
- Time (s) – Recording time point (unit: seconds).
- None –Reaction progress treated by no ions.
- Zn²⁺ – Reaction progress treatmented by Zinc ions.
- Ca²⁺ – Reaction progress treatmented by Calcium ions.
- Zn2+ and Ca2+ – Sequence where Zinc is added first, followed by Calcium.
- Ca2+ and Zn2+ –Sequence where Calcium is added first, followed by Zinc.
- Maximum fusion probability – Measured at the corresponding Zn2+ and Ca2+(unit: Unitless probability (range 0 to 1, where 0 = no fusion, 1 = all vesicles fuse))
SE: Standard Error of the Mean for the Maximum Fusion Probability(unit: Same as the probability (unitless)).
Supplementary Data 8a and b
Description: This file provides the distribution and efficiency changes of Ca²⁺-dependent vesicle fusion with varying protein types, using single-vesicle fusion assay.
Variables
- Time (s) – Recording time point (unit: seconds).
- Ca²⁺-dependent fusion – probability distribution of fusion events per second, recorded over 1 minute in the presence of ions (unitless, Probability scale: Normalized cumulative probability (0-1 )).
- Cumulative probability – Cumulative probability of Ca²⁺-dependent vesicle fusion events by different protein types (unitless).
- Syt1_WT – Wild-type Synaptotagmin-1 reconstituted into vesicles. This is the positive control with all necessary components.
- Syt1_C2AB – synaptotagmin-1 calcium-binding sites mutant reconstituted into vesicles.
- Syt1_4A – synaptotagmin-1 -membrane-insertion mutant reconstituted into vesicles.
- Syt1_4W – synaptotagmin-1 -membrane-insertion mutant reconstituted into vesicles.
Supplementary Data 10
Description: This file provides CD spectrum data for C2AB and its variant proteins.
Variables
- C2AB – Full cytoplasmic domain fragment of synaptotagmin-1.
- C2A – Isolated C2A domain fragment.
- C2B – Isolated C2B domain fragment.
- C2AB – C2AB mutant with disrupted calcium-binding sites.
- C2AB_3M – C2AB mutant with disrupted zinc-binding residues (D261A/E386A/H389A).
- wavelength [nm] – The wavelength of light(unit - Nanometers)
Supplementary Data 11a, b, c and d
Description: This file provides the maximum fusion probability(FRET-based) and efficiency changes of vesicle fusion dependent on different Zn2+ concentration,using lipid mixing assay.
Variables
- Time (s) – Recording time point (unit: seconds).
- Zn2+ concentration(nM) – Treatment with different zinc ion concentrations (unit: nanomolar).
- Syt1_WT – Wild-type Synaptotagmin-1 reconstituted into vesicles.
- Syt1_3M – Syt1 zinc-binding mutant reconstituted into vesicles.
- Syt1_WT(2 mM Mg2+) – Wild-type Synaptotagmin-1 reconstituted into vesicles with 2 mM Mg2+ in solution.
- Syt1_3M(2 mM Mg2+) – Syt1 zinc-binding mutant reconstituted into vesicles with 2 mM Mg2+ in solution.
- Maximum fusion probability – Measured at the corresponding different zinc ion concentrations (unit: Unitless probability (range 0 to 1, where 0 = no fusion, 1 = all vesicles fuse)).
- SE: Standard Error of the Mean for the Maximum Fusion Probability(unit: Same as the probability (unitless)).
Supplementary Data 12a and b
Description: This file provides the maximum fusion probability(FRET-based) and efficiency changes of vesicle fusion dependent on Ca2+ concentration,using lipid mixing assay.
Variables
- Time (s) – Recording time point (unit: seconds).
- Syt1_WT – Wild-type Synaptotagmin-1 reconstituted into vesicles.
- Syt1_3M – Syt1 zinc-binding mutant reconstituted into vesicles.
- Syt1_WT(Ca2+) – Wild-type Synaptotagmin-1 reconstituted into vesicles treatmented with Ca2+.
- Syt1_3M(Ca2+) – Syt1 zinc-binding mutant reconstituted into vesicles treatmented with Ca2+.
- Maximum fusion probability – Measured at the corresponding Ca2+(unit: Unitless probability (range 0 to 1, where 0 = no fusion, 1 = all vesicles fuse)).
- SE: Standard Error of the Mean for the Maximum Fusion Probability(unit: Same as the probability (unitless)).
Supplementary Data 13b
Description: This file provides electrophysiology data from Syt1 cKO hippocampal neurons rescued by Syt1_3M measuring evoked(EPSC) release frequencies and amplitudes under varying Zn²⁺ concentrations.
Variables
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- Evoked EPSC amplitude (pA) – Amplitude of evoked excitatory postsynaptic currents in Syt1 cKO hippocampal neurons rescued by Syt1_3M (unit: picoampere).
Supplementary Data 14b, c, e and f
Description: This file provides electrophysiology data from Syt1 cKO hippocampal neurons rescued by Syt1_WT or Syt1_3M measuring spontaneous(sEPSC) release frequencies and amplitudes under varying Zn²⁺ concentrations.
Variables
- Zn²⁺ concentration (µM) – Treatment with different extracellular zinc ion concentrations (unit: micromolar).
- cKO + Syt1_WT –Syt1 cKO hippocampal neurons rescued by Syt1_WT.
- sEPSC frequency (Hz) – Frequency of spontaneous excitatory postsynaptic currents in Syt1 cKO hippocampal neurons rescued by Syt1_WT neurons(unit: hertz).
- sEPSC amplitude (pA) – Amplitude of spontaneous excitatory postsynaptic currents in Syt1 cKO hippocampal neurons rescued by Syt1_WT neurons(unit: picoampere).
- cKO + Syt1_3M –Syt1 cKO hippocampal neurons rescued by Syt1_3M.
- sEPSC frequency (Hz)– Frequency of spontaneous excitatory postsynaptic currents in Syt1 cKO hippocampal neurons rescued by Syt1_3M neurons(unit: hertz).
- sEPSC amplitude (pA) – Amplitude of spontaneous excitatory postsynaptic currents in Syt1 cKO hippocampal neurons rescued by Syt1_3M neurons (unit: picoampere).
Supplementary Data 15a, b and c
Description: This file provides gel images and precipitation fluorescence spectra related to the study of the binding ability of Ca²⁺ to membranes containing compositions of phosphatidylserine (PS) and phosphatidylinositol (PIP2).
Variables
- Ca²⁺ concentration (µM) – Concentration of Calcium ions in the experimental solution (unit: micromolar).
- C2AB – Cytosolic tandem C2-domains of synaptotagmin-1.
- C2AB– C2AB mutant with disrupted calcium-binding sites.
- C2AB_QQQ – C2AB polybasic region mutant.
- S – Supernatant in fraction.
- P – Pellet in fraction.
- KDa– Kilo-Dalton, measuring protein mass.
- 15% PS + 1% PIP2 – The lipid composition of the vesicles included 15% phosphatidylserine (PS) and 1% phosphatidylinositol 4,5-bisphosphate (PIP₂).
- Intensity (Counts/s/µA) – Fluorescence signal intensity measured at the corresponding peak wavelength (unit: arbitrary units(normalized fluorescence intensity)).
- wavelength [nm] – The wavelength of light(unit - Nanometers).
Normalized P/S – Normalized ratio of protein bound to the pellet fraction versus remaining in the supernatant fraction.
Supplementary Data 16a, b and c
Description: This file provides gel images and precipitation fluorescence spectra related to the study of the binding ability of Zn²⁺ to membranes containing compositions of phosphatidylserine (PS) and phosphatidylinositol (PIP2).
Variables
- Zn²⁺ concentration (µM) – Concentration of zinc ions in the experimental solution (unit: micromolar).
- C2AB – Cytosolic tandem C2-domains of synaptotagmin-1.
- C2AB– C2AB mutant with disrupted calcium-binding sites.
- C2AB_QQQ – C2AB polybasic region mutant.
- S – Supernatant in fraction.
- P – Pellet in fraction.
- KDa– Kilo-Dalton, measuring protein mass.
- 15% PS + 1% PIP2 – The lipid composition of the vesicles included 15% phosphatidylserine (PS) and 1% phosphatidylinositol 4,5-bisphosphate (PIP₂).
- Intensity (Counts/s/µA) – Fluorescence signal intensity measured at the corresponding peak wavelength (unit: arbitrary units(normalized fluorescence intensity)).
- wavelength [nm] – The wavelength of light(unit - Nanometers).
- Normalized P/S – Normalized ratio of protein bound to the pellet fraction versus remaining in the supernatant fraction.
Supplementary Data 17a and b
Description: This file provides the fluorescence response curve and quantitated values for Zn²⁺ influx in hippocampal neurons.
Variables
- Time (s) – Recording time point (unit: seconds).
- Intensity – Fluorescence intensity change of the Zn²⁺ probe in the selected region(Supplementary Data 17a).
- Intensity – Fluorescence values at 0 seconds and 120 seconds(Supplementary Data 17b).
Supplementary Data 18a and b
Description: This file provides the distribution and efficiency changes of Zn²⁺-dependent vesicle fusion with varying protein types, using single-vesicle fusion assay.
Variables
- Time (s) – Recording time point (unit: seconds)
- 500 μM Zn²⁺-dependent fusion probability – probability distribution of fusion events per second, recorded over 1 minute in the presence of 500 μM Zn²⁺ (unitless, Probability scale: Normalized cumulative probability (0-1 )).
- Cumulative probability – Cumulative probability of Zn²⁺-dependent vesicle fusion events by different protein types (unitless).
- Syt1_WT – Wild-type Synaptotagmin-1 reconstituted into vesicles. This is the positive control with all necessary components.
- Syt1_QM – synaptotagmin-1 primary interface with the SNARE complex region mutant reconstituted into vesicles.
- Syt1_QQQ – synaptotagmin-1 polybasic region mutant mutant reconstituted into vesicles.
- Syt1_3M –Syt1 zinc-binding mutant reconstituted into vesicles reconstituted into vesicles.
Access information
Other publicly accessible locations of the data:
- The complex of Syt1/Syt3 and Zn2+ data generated in this study have been deposited in the ModelArchive database under accession codes ma-sle84, ma-36fbl.
Data was derived from the following sources:
- The crystal structure of the C2AB fragment of Syt3 used in this study were obtained from the Protein Data Bank (PDB) with accession code 3HN8.