data_12003 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; ILMV methyl assignments for the neuronal SNARE complex ; _BMRB_accession_number 12003 _BMRB_flat_file_name bmr12003.str _Entry_type original _Submission_date 2016-09-12 _Accession_date 2016-09-13 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details ; Sequence-specific assignment of Methyl Groups from the neuronal SNARE complex using lanthanide-induced pseudocontact shifts ; loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Pan Yun-Zu . . 2 Quade Bradley . . 3 Brewer Kyle . . 4 Graham Bim . . 5 Rizo Josep . . 6 Swarbrick James . . 7 Szabo Monika . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 4 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 111 "13C chemical shifts" 111 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2017-03-07 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 6235 'Backbone 1H, 13C, and 15N Chemical Shift Assignments for the SNARE complex PubMed: 15733924' stop_ _Original_release_date 2016-09-13 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Sequence-specific assignment of methyl groups from the neuronal SNARE complex using lanthanide-induced pseudocontact shifts ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 27988858 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Pan Yun-Zu Z. . 2 Quade Bradley . . 3 Brewer Kyle D. . 4 Szabo Monika . . 5 Swarbrick James D. . 6 Graham Bim . . 7 Rizo Josep . . stop_ _Journal_abbreviation 'J. Biomol. NMR' _Journal_volume 66 _Journal_issue 4 _Journal_ISSN 1573-5001 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 281 _Page_last 293 _Year 2016 _Details . loop_ _Keyword SNAREs 'membrane fusion' 'methyl assignment' 'neurotransmitter release' 'pseudocontact shifts' stop_ save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'neuronal SNARE complex' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'SNAP-25 N-terminal SNARE motif' $SNAP-25N 'SNAP-25 C-terminal SNARE motif' $SNAP-25C 'synaptobrevin-2 SNARE motif' $synaptobrevin_2 'syntaxin-1A SNARE motif' $syntaxin-1A stop_ _System_molecular_weight 32000 _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . loop_ _Biological_function 'neurotransmitter release' stop_ _Database_query_date . _Details 'The neuronal SNARE complex used contained the SNARE motifs of rat syntaxin-1A, SNAP-25A and synaptobrevin 2' save_ ######################## # Monomeric polymers # ######################## save_SNAP-25N _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common SNAP-25N _Molecular_mass . _Mol_thiol_state 'not present' loop_ _Biological_function 'Neurotransmitter release' stop_ _Details ; For assignments purposes, some of the SNARE complexes used in this study contained a D41C mutation in this sequence to attach lanthanide tags. ; ############################## # Polymer residue sequence # ############################## _Residue_count 75 _Mol_residue_sequence ; GSWLEEMQRRADQLADESLE STRRMLQLVEESKDAGIRTL VMLDEQGEQLDRVEEGMNHI NQDMKEAEKNLKDLG ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 8 GLY 2 9 SER 3 10 TRP 4 11 LEU 5 12 GLU 6 13 GLU 7 14 MET 8 15 GLN 9 16 ARG 10 17 ARG 11 18 ALA 12 19 ASP 13 20 GLN 14 21 LEU 15 22 ALA 16 23 ASP 17 24 GLU 18 25 SER 19 26 LEU 20 27 GLU 21 28 SER 22 29 THR 23 30 ARG 24 31 ARG 25 32 MET 26 33 LEU 27 34 GLN 28 35 LEU 29 36 VAL 30 37 GLU 31 38 GLU 32 39 SER 33 40 LYS 34 41 ASP 35 42 ALA 36 43 GLY 37 44 ILE 38 45 ARG 39 46 THR 40 47 LEU 41 48 VAL 42 49 MET 43 50 LEU 44 51 ASP 45 52 GLU 46 53 GLN 47 54 GLY 48 55 GLU 49 56 GLN 50 57 LEU 51 58 ASP 52 59 ARG 53 60 VAL 54 61 GLU 55 62 GLU 56 63 GLY 57 64 MET 58 65 ASN 59 66 HIS 60 67 ILE 61 68 ASN 62 69 GLN 63 70 ASP 64 71 MET 65 72 LYS 66 73 GLU 67 74 ALA 68 75 GLU 69 76 LYS 70 77 ASN 71 78 LEU 72 79 LYS 73 80 ASP 74 81 LEU 75 82 GLY stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ save_SNAP-25C _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common SNAP-25C _Molecular_mass . _Mol_thiol_state 'not present' loop_ _Biological_function 'Neurotransmitter release' stop_ _Details ; For assignments purposes, some of the SNARE complexes used in this study contained a D166C mutation in this sequence to attach lanthanide tags. ; _Residue_count 66 _Mol_residue_sequence ; GSWARENEMDENLEQVSGII GNLRHMALDMGNEIDTQNRQ IDRIMEKADSNKTRIDEANQ RATKML ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 138 GLY 2 139 SER 3 140 TRP 4 141 ALA 5 142 ARG 6 143 GLU 7 144 ASN 8 145 GLU 9 146 MET 10 147 ASP 11 148 GLU 12 149 ASN 13 150 LEU 14 151 GLU 15 152 GLN 16 153 VAL 17 154 SER 18 155 GLY 19 156 ILE 20 157 ILE 21 158 GLY 22 159 ASN 23 160 LEU 24 161 ARG 25 162 HIS 26 163 MET 27 164 ALA 28 165 LEU 29 166 ASP 30 167 MET 31 168 GLY 32 169 ASN 33 170 GLU 34 171 ILE 35 172 ASP 36 173 THR 37 174 GLN 38 175 ASN 39 176 ARG 40 177 GLN 41 178 ILE 42 179 ASP 43 180 ARG 44 181 ILE 45 182 MET 46 183 GLU 47 184 LYS 48 185 ALA 49 186 ASP 50 187 SER 51 188 ASN 52 189 LYS 53 190 THR 54 191 ARG 55 192 ILE 56 193 ASP 57 194 GLU 58 195 ALA 59 196 ASN 60 197 GLN 61 198 ARG 62 199 ALA 63 200 THR 64 201 LYS 65 202 MET 66 203 LEU stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ save_synaptobrevin_2 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common synaptobrevin_2 _Molecular_mass . _Mol_thiol_state 'not present' loop_ _Biological_function 'Neurotransmitter release' stop_ _Details . _Residue_count 67 _Mol_residue_sequence ; GSNRRLQQTQAQVDEVVDIM RVNVDKVLERDQKLSELDDR ADALQAGASQFETSAAKLKR KYWWKNL ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 27 GLY 2 28 SER 3 29 ASN 4 30 ARG 5 31 ARG 6 32 LEU 7 33 GLN 8 34 GLN 9 35 THR 10 36 GLN 11 37 ALA 12 38 GLN 13 39 VAL 14 40 ASP 15 41 GLU 16 42 VAL 17 43 VAL 18 44 ASP 19 45 ILE 20 46 MET 21 47 ARG 22 48 VAL 23 49 ASN 24 50 VAL 25 51 ASP 26 52 LYS 27 53 VAL 28 54 LEU 29 55 GLU 30 56 ARG 31 57 ASP 32 58 GLN 33 59 LYS 34 60 LEU 35 61 SER 36 62 GLU 37 63 LEU 38 64 ASP 39 65 ASP 40 66 ARG 41 67 ALA 42 68 ASP 43 69 ALA 44 70 LEU 45 71 GLN 46 72 ALA 47 73 GLY 48 74 ALA 49 75 SER 50 76 GLN 51 77 PHE 52 78 GLU 53 79 THR 54 80 SER 55 81 ALA 56 82 ALA 57 83 LYS 58 84 LEU 59 85 LYS 60 86 ARG 61 87 LYS 62 88 TYR 63 89 TRP 64 90 TRP 65 91 LYS 66 92 ASN 67 93 LEU stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ save_syntaxin-1A _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common syntaxin-1A _Molecular_mass . _Mol_thiol_state 'not present' loop_ _Biological_function 'Neurotransmitter release' stop_ _Details ; For assignments purposes, some of the SNARE complexes used in this study contained a D214C mutation in this sequence to attach lanthanide tags. ; _Residue_count 65 _Mol_residue_sequence ; GSALSEIETRHSEIIKLENS IRELHDMFMDMAMLVESQGE MIDRIEYNVEHAVDYVERAV SDTKK ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 189 GLY 2 190 SER 3 191 ALA 4 192 LEU 5 193 SER 6 194 GLU 7 195 ILE 8 196 GLU 9 197 THR 10 198 ARG 11 199 HIS 12 200 SER 13 201 GLU 14 202 ILE 15 203 ILE 16 204 LYS 17 205 LEU 18 206 GLU 19 207 ASN 20 208 SER 21 209 ILE 22 210 ARG 23 211 GLU 24 212 LEU 25 213 HIS 26 214 ASP 27 215 MET 28 216 PHE 29 217 MET 30 218 ASP 31 219 MET 32 220 ALA 33 221 MET 34 222 LEU 35 223 VAL 36 224 GLU 37 225 SER 38 226 GLN 39 227 GLY 40 228 GLU 41 229 MET 42 230 ILE 43 231 ASP 44 232 ARG 45 233 ILE 46 234 GLU 47 235 TYR 48 236 ASN 49 237 VAL 50 238 GLU 51 239 HIS 52 240 ALA 53 241 VAL 54 242 ASP 55 243 TYR 56 244 VAL 57 245 GLU 58 246 ARG 59 247 ALA 60 248 VAL 61 249 SER 62 250 ASP 63 251 THR 64 252 LYS 65 253 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $SNAP-25N Rat 10116 Eukaryota Metazoa Rattus norvegicus $SNAP-25C Rat 10116 Eukaryota Metazoa Rattus norvegicus $synaptobrevin_2 Rat 10116 Eukaryota Metazoa Rattus norvegicus $syntaxin-1A Rat 10116 Eukaryota Metazoa Rattus norvegicus stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Variant _Vector_name $SNAP-25N 'recombinant technology' . Escherichia coli BL21 DE3 pET28a $SNAP-25C 'recombinant technology' . Escherichia coli BL21 DE3 pET28a $synaptobrevin_2 'recombinant technology' . Escherichia coli BL21 DE3 pET28a $syntaxin-1A 'recombinant technology' . Escherichia coli BL21 DE3 pET28a stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details ; We used multiple samples that were essentially identical to sample 1 but contained lanthanide tags placed at different positions and were isotopically labeled at different SNARE motifs. ; loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $SNAP-25N 30 uM '[ILMV-13CH3; U-2H]' $SNAP-25C 30 uM 'natural abundance' $synaptobrevin_2 30 uM 'natural abundance' $syntaxin-1A 30 uM 'natural abundance' 'sodium chloride' 200 mM 'natural abundance' TRIS 20 mM 'natural abundance' H2O 90 % 'natural abundance' D2O 10 % [U-2H] stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details ; We used multiple samples that were essentially identical to sample 1 but contained lanthanide tags placed at different positions and were isotopically labeled at different SNARE motifs. ; loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $SNAP-25N 30 uM 'natural abundance' $SNAP-25C 30 uM '[ILMV-13CH3; U-2H]' $synaptobrevin_2 30 uM 'natural abundance' $syntaxin-1A 30 uM 'natural abundance' 'sodium chloride' 200 mM 'natural abundance' TRIS 20 mM 'natural abundance' H2O 90 % 'natural abundance' D2O 10 % [U-2H] stop_ save_ save_sample_3 _Saveframe_category sample _Sample_type solution _Details ; We used multiple samples that were essentially identical to sample 1 but contained lanthanide tags placed at different positions and were isotopically labeled at different SNARE motifs. ; loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $SNAP-25N 30 uM 'natural abundance' $SNAP-25C 30 uM 'natural abundance' $synaptobrevin_2 30 uM '[ILMV-13CH3; U-2H]' $syntaxin-1A 30 uM 'natural abundance' 'sodium chloride' 200 mM 'natural abundance' TRIS 20 mM 'natural abundance' H2O 90 % 'natural abundance' D2O 10 % [U-2H] stop_ save_ save_sample_4 _Saveframe_category sample _Sample_type solution _Details ; We used multiple samples that were essentially identical to sample 1 but contained lanthanide tags placed at different positions and were isotopically labeled at different SNARE motifs. ; loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $SNAP-25N 30 uM 'natural abundance' $SNAP-25C 30 uM 'natural abundance' $synaptobrevin_2 30 uM 'natural abundance' $syntaxin-1A 30 uM '[ILMV-13CH3; U-2H]' 'sodium chloride' 200 mM 'natural abundance' TRIS 20 mM 'natural abundance' H2O 90 % 'natural abundance' D2O 10 % [U-2H] stop_ save_ ############################ # Computer software used # ############################ save_NMRPipe _Saveframe_category software _Name NMRPipe _Version . loop_ _Vendor _Address _Electronic_address 'Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax' . . stop_ loop_ _Task processing stop_ _Details . save_ save_NMRView _Saveframe_category software _Name NMRView _Version . loop_ _Vendor _Address _Electronic_address 'Johnson, One Moon Scientific' . . stop_ loop_ _Task 'data analysis' stop_ _Details . save_ save_NUMBAT _Saveframe_category software _Name NUMBAT _Version . loop_ _Vendor _Address _Electronic_address 'Christophe Schmitz' . . stop_ loop_ _Task 'PCS analysis' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Agilent _Model DD2 _Field_strength 800 _Details . save_ save_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Agilent _Model DD2 _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-13C_HMQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HMQC' _Sample_label $sample_1 save_ save_2D_1H-13C_HMQC_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HMQC' _Sample_label $sample_2 save_ save_2D_1H-13C_HMQC_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HMQC' _Sample_label $sample_3 save_ save_2D_1H-13C_HMQC_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HMQC' _Sample_label $sample_4 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 220 . mM pH 7.4 . pH pressure 1 . atm temperature 298 . K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference_1 _Saveframe_category chemical_shift_reference _Details . loop_ _Mol_common_name _Atom_type _Atom_isotope_number _Atom_group _Chem_shift_units _Chem_shift_value _Reference_method _Reference_type _External_reference_sample_geometry _External_reference_location _External_reference_axis _Indirect_shift_ratio DSS C 13 'methyl protons' ppm 0.00 na indirect . . . 0.251449530 DSS H 1 'methyl protons' ppm 0.00 internal direct . . . 1.000000000 stop_ save_ ################################### # Assigned chemical shift lists # ################################### ################################################################### # Chemical Shift Ambiguity Index Value Definitions # # # # The values other than 1 are used for those atoms with different # # chemical shifts that cannot be assigned to stereospecific atoms # # or to specific residues or chains. # # # # Index Value Definition # # # # 1 Unique (including isolated methyl protons, # # geminal atoms, and geminal methyl # # groups with identical chemical shifts) # # (e.g. ILE HD11, HD12, HD13 protons) # # 2 Ambiguity of geminal atoms or geminal methyl # # proton groups (e.g. ASP HB2 and HB3 # # protons, LEU CD1 and CD2 carbons, or # # LEU HD11, HD12, HD13 and HD21, HD22, # # HD23 methyl protons) # # 3 Aromatic atoms on opposite sides of # # symmetrical rings (e.g. TYR HE1 and HE2 # # protons) # # 4 Intraresidue ambiguities (e.g. LYS HG and # # HD protons or TRP HZ2 and HZ3 protons) # # 5 Interresidue ambiguities (LYS 12 vs. LYS 27) # # 6 Intermolecular ambiguities (e.g. ASP 31 CA # # in monomer 1 and ASP 31 CA in monomer 2 # # of an asymmetrical homodimer, duplex # # DNA assignments, or other assignments # # that may apply to atoms in one or more # # molecule in the molecular assembly) # # 9 Ambiguous, specific ambiguity not defined # # # ################################################################### save_assigned_chem_shift_list_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-13C HMQC' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name 'SNAP-25 N-terminal SNARE motif' _Text_data_format . _Text_data . loop_ _Atom_shift_assign_ID _Residue_author_seq_code _Residue_seq_code _Residue_label _Atom_name _Atom_type _Chem_shift_value _Chem_shift_value_error _Chem_shift_ambiguity_code 1 11 4 LEU HD1 H 0.855 0.005 1 2 11 4 LEU HD2 H 0.760 0.005 1 3 11 4 LEU CD1 C 24.19 0.03 1 4 11 4 LEU CD2 C 24.09 0.03 1 5 14 7 MET HE H 2.008 0.005 1 6 14 7 MET CE C 17.52 0.03 1 7 21 14 LEU HD1 H 0.633 0.005 1 8 21 14 LEU HD2 H 0.308 0.005 1 9 21 14 LEU CD1 C 25.74 0.03 1 10 21 14 LEU CD2 C 21.88 0.03 1 11 26 19 LEU HD1 H 0.842 0.005 1 12 26 19 LEU HD2 H 0.983 0.005 1 13 26 19 LEU CD1 C 22.37 0.03 1 14 26 19 LEU CD2 C 25.76 0.03 1 15 32 25 MET HE H 2.131 0.005 1 16 32 25 MET CE C 20.04 0.03 1 17 33 26 LEU HD1 H 0.972 0.005 1 18 33 26 LEU HD2 H 0.852 0.005 1 19 33 26 LEU CD1 C 25.36 0.03 1 20 33 26 LEU CD2 C 22.61 0.03 1 21 35 28 LEU HD1 H 0.699 0.005 1 22 35 28 LEU HD2 H 0.238 0.005 1 23 35 28 LEU CD1 C 25.66 0.03 1 24 35 28 LEU CD2 C 21.97 0.03 1 25 36 29 VAL HG1 H 1.091 0.005 1 26 36 29 VAL HG2 H 1.364 0.005 1 27 36 29 VAL CG1 C 23.30 0.03 1 28 36 29 VAL CG2 C 21.08 0.03 1 29 44 37 ILE HD1 H 0.890 0.005 1 30 44 37 ILE CD1 C 13.38 0.03 1 31 47 40 LEU HD1 H 0.787 0.005 1 32 47 40 LEU HD2 H 0.790 0.005 1 33 47 40 LEU CD1 C 24.04 0.03 1 34 47 40 LEU CD2 C 24.32 0.03 1 35 48 41 VAL HG1 H 0.951 0.005 1 36 48 41 VAL HG2 H 1.105 0.005 1 37 48 41 VAL CG1 C 21.01 0.03 1 38 48 41 VAL CG2 C 22.61 0.03 1 39 49 42 MET HE H 2.19 0.005 1 40 49 42 MET CE C 17.17 0.03 1 41 50 43 LEU HD1 H 0.766 0.005 1 42 50 43 LEU HD2 H 0.690 0.005 1 43 50 43 LEU CD1 C 25.96 0.03 1 44 50 43 LEU CD2 C 22.12 0.03 1 45 57 50 LEU HD1 H 0.773 0.005 1 46 57 50 LEU HD2 H 0.611 0.005 1 47 57 50 LEU CD1 C 24.98 0.03 1 48 57 50 LEU CD2 C 21.37 0.03 1 49 60 53 VAL HG1 H 0.965 0.005 1 50 60 53 VAL HG2 H 1.051 0.005 1 51 60 53 VAL CG1 C 22.51 0.03 1 52 60 53 VAL CG2 C 23.00 0.03 1 53 64 57 MET HE H 1.974 0.005 1 54 64 57 MET CE C 20.02 0.03 1 55 67 60 ILE HD1 H 0.837 0.005 1 56 67 60 ILE CD1 C 14.51 0.03 1 57 71 64 MET HE H 1.370 0.005 1 58 71 64 MET CE C 14.52 0.03 1 59 78 71 LEU HD1 H 0.983 0.005 1 60 78 71 LEU HD2 H 0.914 0.005 1 61 78 71 LEU CD1 C 25.76 0.03 1 62 78 71 LEU CD2 C 25.46 0.03 1 63 81 74 LEU HD1 H 0.772 0.005 1 64 81 74 LEU HD2 H 0.626 0.005 1 65 81 74 LEU CD1 C 25.38 0.03 1 66 81 74 LEU CD2 C 23.14 0.03 1 stop_ save_ save_assigned_chem_shift_list_2 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-13C HMQC' stop_ loop_ _Sample_label $sample_2 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name 'SNAP-25 C-terminal SNARE motif' _Text_data_format . _Text_data . loop_ _Atom_shift_assign_ID _Residue_author_seq_code _Residue_seq_code _Residue_label _Atom_name _Atom_type _Chem_shift_value _Chem_shift_value_error _Chem_shift_ambiguity_code 1 146 9 MET HE H 2.038 0.005 1 2 146 9 MET CE C 17.09 0.03 1 3 150 13 LEU HD1 H 0.860 0.005 1 4 150 13 LEU HD2 H 0.773 0.005 1 5 150 13 LEU CD1 C 25.41 0.03 1 6 150 13 LEU CD2 C 22.98 0.03 1 7 153 16 VAL HG1 H 0.821 0.005 1 8 153 16 VAL HG2 H 1.001 0.005 1 9 153 16 VAL CG1 C 21.37 0.03 1 10 153 16 VAL CG2 C 23.94 0.03 1 11 156 19 ILE HD1 H 0.804 0.005 1 12 156 19 ILE CD1 C 13.57 0.03 1 13 157 20 ILE HD1 H 0.673 0.005 1 14 157 20 ILE CD1 C 11.96 0.03 1 15 160 23 LEU HD1 H 0.719 0.005 1 16 160 23 LEU HD2 H 0.192 0.005 1 17 160 23 LEU CD1 C 25.60 0.03 1 18 160 23 LEU CD2 C 20.05 0.03 1 19 163 26 MET HE H 2.139 0.005 1 20 163 26 MET CE C 17.97 0.03 1 21 165 28 LEU HD1 H 0.849 0.005 1 22 165 28 LEU HD2 H 0.908 0.005 1 23 165 28 LEU CD1 C 25.49 0.03 1 24 165 28 LEU CD2 C 22.76 0.03 1 25 167 30 MET HE H 2.137 0.005 1 26 167 30 MET CE C 16.84 0.03 1 27 171 34 ILE HD1 H 0.639 0.005 1 28 171 34 ILE CD1 C 13.78 0.03 1 29 178 41 ILE HD1 H 0.908 0.005 1 30 178 41 ILE CD1 C 14.33 0.03 1 31 181 44 ILE HD1 H 0.864 0.005 1 32 181 44 ILE CD1 C 15.18 0.03 1 33 182 45 MET HE H 2.137 0.005 1 34 182 45 MET CE C 17.17 0.03 1 35 192 55 ILE HD1 H 0.716 0.005 1 36 192 55 ILE CD1 C 13.87 0.03 1 37 202 65 MET HE H 1.866 0.005 1 38 202 65 MET CE C 16.74 0.03 1 39 203 66 LEU HD1 H 0.930 0.005 1 40 203 66 LEU HD2 H 0.817 0.005 1 41 203 66 LEU CD1 C 25.71 0.03 1 42 203 66 LEU CD2 C 23.31 0.03 1 stop_ save_ save_assigned_chem_shift_list_3 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-13C HMQC' stop_ loop_ _Sample_label $sample_3 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name 'synaptobrevin-2 SNARE motif' _Text_data_format . _Text_data . loop_ _Atom_shift_assign_ID _Residue_author_seq_code _Residue_seq_code _Residue_label _Atom_name _Atom_type _Chem_shift_value _Chem_shift_value_error _Chem_shift_ambiguity_code 1 32 6 LEU HD1 H 0.941 0.005 1 2 32 6 LEU HD2 H 0.919 0.005 1 3 32 6 LEU CD1 C 25.95 0.03 1 4 32 6 LEU CD2 C 25.25 0.03 1 5 39 13 VAL HG1 H 0.897 0.005 5 6 39 13 VAL HG2 H 1.011 0.005 5 7 39 13 VAL CG1 C 21.91 0.03 1 8 39 13 VAL CG2 C 24.23 0.03 1 9 42 16 VAL HG1 H 1.156 0.005 1 10 42 16 VAL HG2 H 1.188 0.005 1 11 42 16 VAL CG1 C 24.22 0.03 1 12 42 16 VAL CG2 C 23.39 0.03 1 13 43 17 VAL HG1 H 0.902 0.005 5 14 43 17 VAL HG2 H 1.092 0.005 5 15 43 17 VAL CG1 C 21.06 0.03 1 16 43 17 VAL CG2 C 24.34 0.03 1 17 45 19 ILE HD1 H 0.876 0.005 1 18 45 19 ILE CD1 C 14.30 0.03 1 19 46 20 MET HE H 1.030 0.005 1 20 46 20 MET CE C 14.87 0.03 1 21 48 22 VAL HG1 H 1.028 0.005 1 22 48 22 VAL HG2 H 1.105 0.005 1 23 48 22 VAL CG1 C 21.60 0.03 1 24 48 22 VAL CG2 C 22.44 0.03 1 25 50 24 VAL HG1 H 0.902 0.005 1 26 50 24 VAL HG2 H 1.003 0.005 5 27 50 24 VAL CG1 C 21.06 0.03 1 28 50 24 VAL CG2 C 22.54 0.03 1 29 53 27 VAL HG1 H 1.284 0.005 1 30 53 27 VAL HG2 H 1.193 0.005 1 31 53 27 VAL CG1 C 22.19 0.03 1 32 53 27 VAL CG2 C 25.72 0.03 1 33 54 28 LEU HD1 H 0.876 0.005 1 34 54 28 LEU HD2 H 0.867 0.005 1 35 54 28 LEU CD1 C 25.14 0.03 1 36 54 28 LEU CD2 C 22.25 0.03 1 37 60 34 LEU HD1 H 0.892 0.005 1 38 60 34 LEU HD2 H 0.746 0.005 1 39 60 34 LEU CD1 C 27.13 0.03 1 40 60 34 LEU CD2 C 22.83 0.03 1 41 63 37 LEU HD1 H 0.900 0.005 1 42 63 37 LEU HD2 H 0.925 0.005 1 43 63 37 LEU CD1 C 22.03 0.03 1 44 63 37 LEU CD2 C 26.22 0.03 1 45 70 44 LEU HD1 H 0.947 0.005 1 46 70 44 LEU HD2 H 0.925 0.005 1 47 70 44 LEU CD1 C 24.79 0.03 1 48 70 44 LEU CD2 C 26.22 0.03 1 49 84 58 LEU HD1 H 0.880 0.005 1 50 84 58 LEU HD2 H 0.934 0.005 1 51 84 58 LEU CD1 C 25.40 0.03 1 52 84 58 LEU CD2 C 24.50 0.03 1 53 93 67 LEU HD1 H 0.875 0.005 1 54 93 67 LEU HD2 H 0.834 0.005 1 55 93 67 LEU CD1 C 25.14 0.03 1 56 93 67 LEU CD2 C 23.39 0.03 1 stop_ loop_ _Atom_shift_assign_ID_ambiguity 5 13 '5,13' '5,13' '6,14,26' '6,14,26' '6,15,26' stop_ save_ save_assigned_chem_shift_list_4 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-13C HMQC' stop_ loop_ _Sample_label $sample_4 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name 'syntaxin-1A SNARE motif' _Text_data_format . _Text_data . loop_ _Atom_shift_assign_ID _Residue_author_seq_code _Residue_seq_code _Residue_label _Atom_name _Atom_type _Chem_shift_value _Chem_shift_value_error _Chem_shift_ambiguity_code 1 192 4 LEU HD1 H 0.890 0.005 1 2 192 4 LEU HD2 H 0.891 0.005 1 3 192 4 LEU CD1 C 23.94 0.03 1 4 192 4 LEU CD2 C 24.62 0.03 1 5 195 7 ILE HD1 H 0.890 0.005 1 6 195 7 ILE CD1 C 13.15 0.03 1 7 202 14 ILE HD1 H 0.866 0.005 1 8 202 14 ILE CD1 C 13.21 0.03 1 9 203 15 ILE HD1 H 0.624 0.005 1 10 203 15 ILE CD1 C 9.28 0.03 1 11 205 17 LEU HD1 H 0.801 0.005 1 12 205 17 LEU HD2 H 0.799 0.005 1 13 205 17 LEU CD1 C 23.26 0.03 1 14 205 17 LEU CD2 C 26.61 0.03 1 15 209 21 ILE HD1 H 0.730 0.005 1 16 209 21 ILE CD1 C 13.86 0.03 1 17 212 24 LEU HD1 H 1.143 0.005 1 18 212 24 LEU HD2 H 1.064 0.005 1 19 212 24 LEU CD1 C 23.99 0.03 1 20 212 24 LEU CD2 C 26.31 0.03 1 21 215 27 MET HE H 2.133 0.005 1 22 215 27 MET CE C 18.21 0.03 1 23 217 29 MET HE H 1.928 0.005 1 24 217 29 MET CE C 16.96 0.03 1 25 219 31 MET HE H 1.805 0.005 1 26 219 31 MET CE C 17.31 0.03 1 27 221 33 MET HE H 2.117 0.005 1 28 221 33 MET CE C 16.93 0.03 1 29 222 34 LEU HD1 H 0.807 0.005 1 30 222 34 LEU HD2 H 0.859 0.005 1 31 222 34 LEU CD1 C 24.95 0.03 1 32 222 34 LEU CD2 C 23.41 0.03 1 33 223 35 VAL HG1 H 1.048 0.005 1 34 223 35 VAL HG2 H 0.993 0.005 1 35 223 35 VAL CG1 C 21.88 0.03 1 36 223 35 VAL CG2 C 24.40 0.03 1 37 229 41 MET HE H 2.201 0.005 1 38 229 41 MET CE C 17.46 0.03 1 39 230 42 ILE HD1 H 0.797 0.005 1 40 230 42 ILE CD1 C 13.50 0.03 1 41 233 45 ILE HD1 H 0.826 0.005 1 42 233 45 ILE CD1 C 14.62 0.03 1 43 237 49 VAL HG1 H 1.087 0.005 1 44 237 49 VAL HG2 H 1.123 0.005 1 45 237 49 VAL CG1 C 21.28 0.03 1 46 237 49 VAL CG2 C 25.08 0.03 1 47 241 53 VAL HG1 H 1.116 0.005 1 48 241 53 VAL HG2 H 1.116 0.005 1 49 241 53 VAL CG1 C 23.35 0.03 1 50 241 53 VAL CG2 C 23.00 0.03 1 51 244 56 VAL HG1 H 0.687 0.005 1 52 244 56 VAL HG2 H 1.246 0.005 1 53 244 56 VAL CG1 C 20.71 0.03 1 54 244 56 VAL CG2 C 24.13 0.03 1 55 248 60 VAL HG1 H 0.950 0.005 1 56 248 60 VAL HG2 H 1.129 0.005 1 57 248 60 VAL CG1 C 21.52 0.03 1 58 248 60 VAL CG2 C 23.65 0.03 1 stop_ save_