data_36146 ####################### # Entry information # ####################### save_entry_information _Entry.Sf_category entry_information _Entry.Sf_framecode entry_information _Entry.ID 36146 _Entry.Title ; mBjAMP1 structure ; _Entry.Type macromolecule _Entry.Version_type original _Entry.Submission_date 2017-12-28 _Entry.Accession_date 2018-06-27 _Entry.Last_release_date 2018-06-27 _Entry.Original_release_date 2018-06-27 _Entry.Origination author _Entry.Format_name . _Entry.NMR_STAR_version 3.2.0.16 _Entry.Original_NMR_STAR_version 3.2.0.16 _Entry.Experimental_method NMR _Entry.Experimental_method_subtype SOLUTION _Entry.Source_data_format . _Entry.Source_data_format_version . _Entry.Generated_software_name . _Entry.Generated_software_version . _Entry.Generated_software_ID . _Entry.Generated_software_label . _Entry.Generated_date . _Entry.DOI . _Entry.UUID . _Entry.Related_coordinate_file_name . _Entry.Details . _Entry.BMRB_internal_directory_name . loop_ _Entry_experimental_methods.ID _Entry_experimental_methods.Method _Entry_experimental_methods.Subtype _Entry_experimental_methods.Entry_ID 1 'SOLUTION NMR' 'SOLUTION NMR' 36146 stop_ loop_ _Entry_author.Ordinal _Entry_author.Given_name _Entry_author.Family_name _Entry_author.First_initial _Entry_author.Middle_initials _Entry_author.Family_title _Entry_author.ORCID _Entry_author.Entry_ID 1 J. Nam J. Y. . . 36146 2 C. Lee C. W. . . 36146 stop_ loop_ _Struct_keywords.Keywords _Struct_keywords.Text _Struct_keywords.Entry_ID ANTIBIOTIC . 36146 'Antimicrobial peptides' . 36146 'Cysteine rich peptides' . 36146 Folding . 36146 stop_ loop_ _Data_set.Type _Data_set.Count _Data_set.Entry_ID assigned_chemical_shifts 1 36146 stop_ loop_ _Datum.Type _Datum.Count _Datum.Entry_ID '1H chemical shifts' 138 36146 stop_ loop_ _Release.Release_number _Release.Format_type _Release.Format_version _Release.Date _Release.Submission_date _Release.Type _Release.Author _Release.Detail _Release.Entry_ID 2 . . 2019-06-21 2017-12-28 update BMRB 'update entry citation' 36146 1 . . 2018-12-18 2017-12-28 original author 'original release' 36146 stop_ loop_ _Related_entries.Database_name _Related_entries.Database_accession_code _Related_entries.Relationship _Related_entries.Entry_ID PDB 5Z1Y . 36146 stop_ save_ ############### # Citations # ############### save_citation_1 _Citation.Sf_category citations _Citation.Sf_framecode citation_1 _Citation.Entry_ID 36146 _Citation.ID 1 _Citation.Class 'entry citation' _Citation.CAS_abstract_code . _Citation.MEDLINE_UI_code . _Citation.DOI 10.1021/acs.jmedchem.8b01135 _Citation.PubMed_ID 30475621 _Citation.Full_citation . _Citation.Title ; Structural and functional assessment of mBjAMP1, an antimicrobial peptide from Branchiostoma japonicum, revealed a novel alpha-hairpinin-like scaffold with membrane permeable and DNA binding activity. ; _Citation.Status published _Citation.Type journal _Citation.Journal_abbrev 'J. Med. Chem.' _Citation.Journal_name_full 'Journal of medicinal chemistry' _Citation.Journal_volume 61 _Citation.Journal_issue 24 _Citation.Journal_ASTM . _Citation.Journal_ISSN 0022-2623 _Citation.Journal_CSD 0353 _Citation.Book_title . _Citation.Book_chapter_title . _Citation.Book_volume . _Citation.Book_series . _Citation.Book_publisher . _Citation.Book_publisher_city . _Citation.Book_ISBN . _Citation.Conference_title . _Citation.Conference_site . _Citation.Conference_state_province . _Citation.Conference_country . _Citation.Conference_start_date . _Citation.Conference_end_date . _Citation.Conference_abstract_number . _Citation.Thesis_institution . _Citation.Thesis_institution_city . _Citation.Thesis_institution_country . _Citation.WWW_URL . _Citation.Page_first 11101 _Citation.Page_last 11113 _Citation.Year 2018 _Citation.Details . loop_ _Citation_author.Ordinal _Citation_author.Given_name _Citation_author.Family_name _Citation_author.First_initial _Citation_author.Middle_initials _Citation_author.Family_title _Citation_author.ORCID _Citation_author.Entry_ID _Citation_author.Citation_ID 1 Jiyong Nam J. Y. . . 36146 1 2 Hyosuk Yun . . . . 36146 1 3 Ganesan Rajasekaran . . . . 36146 1 4 'S. Dinesh' Kumar . . . . 36146 1 5 'Jae Il' Kim . . . . 36146 1 6 'Hye Jung' Min . . . . 36146 1 7 'Song Yub' Shin . . . . 36146 1 8 'Chul Won' Lee C. W. . . 36146 1 stop_ save_ ############################################# # Molecular system (assembly) description # ############################################# save_assembly _Assembly.Sf_category assembly _Assembly.Sf_framecode assembly _Assembly.Entry_ID 36146 _Assembly.ID 1 _Assembly.Name 'mBjAMP1 peptide' _Assembly.BMRB_code . _Assembly.Number_of_components . _Assembly.Organic_ligands . _Assembly.Metal_ions . _Assembly.Non_standard_bonds . _Assembly.Ambiguous_conformational_states . _Assembly.Ambiguous_chem_comp_sites . _Assembly.Molecules_in_chemical_exchange . _Assembly.Paramagnetic no _Assembly.Thiol_state . _Assembly.Molecular_mass . _Assembly.Enzyme_commission_number . _Assembly.Details . _Assembly.DB_query_date . _Assembly.DB_query_revised_last_date . loop_ _Entity_assembly.ID _Entity_assembly.Entity_assembly_name _Entity_assembly.Entity_ID _Entity_assembly.Entity_label _Entity_assembly.Asym_ID _Entity_assembly.PDB_chain_ID _Entity_assembly.Experimental_data_reported _Entity_assembly.Physical_state _Entity_assembly.Conformational_isomer _Entity_assembly.Chemical_exchange_state _Entity_assembly.Magnetic_equivalence_group_code _Entity_assembly.Role _Entity_assembly.Details _Entity_assembly.Entry_ID _Entity_assembly.Assembly_ID 1 entity_1 1 $entity_1 A A yes . . . . . . 36146 1 stop_ loop_ _Bond.ID _Bond.Type _Bond.Value_order _Bond.Assembly_atom_ID_1 _Bond.Entity_assembly_ID_1 _Bond.Entity_assembly_name_1 _Bond.Entity_ID_1 _Bond.Comp_ID_1 _Bond.Comp_index_ID_1 _Bond.Seq_ID_1 _Bond.Atom_ID_1 _Bond.Assembly_atom_ID_2 _Bond.Entity_assembly_ID_2 _Bond.Entity_assembly_name_2 _Bond.Entity_ID_2 _Bond.Comp_ID_2 _Bond.Comp_index_ID_2 _Bond.Seq_ID_2 _Bond.Atom_ID_2 _Bond.Auth_entity_assembly_ID_1 _Bond.Auth_entity_assembly_name_1 _Bond.Auth_asym_ID_1 _Bond.Auth_seq_ID_1 _Bond.Auth_comp_ID_1 _Bond.Auth_atom_ID_1 _Bond.Auth_entity_assembly_ID_2 _Bond.Auth_entity_assembly_name_2 _Bond.Auth_asym_ID_2 _Bond.Auth_seq_ID_2 _Bond.Auth_comp_ID_2 _Bond.Auth_atom_ID_2 _Bond.Entry_ID _Bond.Assembly_ID 1 disulfide single . 1 entity_1 1 CYS 3 3 SG . 1 entity_1 1 CYS 15 15 SG . . . 3 CYS . . . . 15 CYS . 36146 1 stop_ loop_ _Entity_deleted_atom.ID _Entity_deleted_atom.Entity_atom_list_ID _Entity_deleted_atom.Entity_assembly_ID _Entity_deleted_atom.Entity_ID _Entity_deleted_atom.Comp_ID _Entity_deleted_atom.Comp_index_ID _Entity_deleted_atom.Seq_ID _Entity_deleted_atom.Atom_ID _Entity_deleted_atom.Auth_entity_assembly_ID _Entity_deleted_atom.Auth_seq_ID _Entity_deleted_atom.Auth_comp_ID _Entity_deleted_atom.Auth_atom_ID _Entity_deleted_atom.Entry_ID _Entity_deleted_atom.Assembly_ID 1 . 1 1 CYS 3 3 HG . 3 . . 36146 1 2 . 1 1 CYS 15 15 HG . 15 . . 36146 1 stop_ save_ #################################### # Biological polymers and ligands # #################################### save_entity_1 _Entity.Sf_category entity _Entity.Sf_framecode entity_1 _Entity.Entry_ID 36146 _Entity.ID 1 _Entity.BMRB_code . _Entity.Name 'mBjAMP1 peptide' _Entity.Type polymer _Entity.Polymer_common_type . _Entity.Polymer_type polypeptide(L) _Entity.Polymer_type_details . _Entity.Polymer_strand_ID A _Entity.Polymer_seq_one_letter_code_can . _Entity.Polymer_seq_one_letter_code ; NLCASLRARHTIPQCRKFGR R ; _Entity.Target_identifier . _Entity.Polymer_author_defined_seq . _Entity.Polymer_author_seq_details . _Entity.Ambiguous_conformational_states . _Entity.Ambiguous_chem_comp_sites . _Entity.Nstd_monomer no _Entity.Nstd_chirality . _Entity.Nstd_linkage no _Entity.Nonpolymer_comp_ID . _Entity.Nonpolymer_comp_label . _Entity.Number_of_monomers 21 _Entity.Number_of_nonpolymer_components . _Entity.Paramagnetic no _Entity.Thiol_state 'all disulfide bound' _Entity.Src_method syn _Entity.Parent_entity_ID . _Entity.Fragment . _Entity.Mutation . _Entity.EC_number . _Entity.Calc_isoelectric_point . _Entity.Formula_weight 2490.979 _Entity.Formula_weight_exptl . _Entity.Formula_weight_exptl_meth . _Entity.Details . _Entity.DB_query_date . _Entity.DB_query_revised_last_date . loop_ _Entity_comp_index.ID _Entity_comp_index.Auth_seq_ID _Entity_comp_index.Comp_ID _Entity_comp_index.Comp_label _Entity_comp_index.Entry_ID _Entity_comp_index.Entity_ID 1 1 ASN . 36146 1 2 2 LEU . 36146 1 3 3 CYS . 36146 1 4 4 ALA . 36146 1 5 5 SER . 36146 1 6 6 LEU . 36146 1 7 7 ARG . 36146 1 8 8 ALA . 36146 1 9 9 ARG . 36146 1 10 10 HIS . 36146 1 11 11 THR . 36146 1 12 12 ILE . 36146 1 13 13 PRO . 36146 1 14 14 GLN . 36146 1 15 15 CYS . 36146 1 16 16 ARG . 36146 1 17 17 LYS . 36146 1 18 18 PHE . 36146 1 19 19 GLY . 36146 1 20 20 ARG . 36146 1 21 21 ARG . 36146 1 stop_ loop_ _Entity_poly_seq.Hetero _Entity_poly_seq.Mon_ID _Entity_poly_seq.Num _Entity_poly_seq.Comp_index_ID _Entity_poly_seq.Entry_ID _Entity_poly_seq.Entity_ID . ASN 1 1 36146 1 . LEU 2 2 36146 1 . CYS 3 3 36146 1 . ALA 4 4 36146 1 . SER 5 5 36146 1 . LEU 6 6 36146 1 . ARG 7 7 36146 1 . ALA 8 8 36146 1 . ARG 9 9 36146 1 . HIS 10 10 36146 1 . THR 11 11 36146 1 . ILE 12 12 36146 1 . PRO 13 13 36146 1 . GLN 14 14 36146 1 . CYS 15 15 36146 1 . ARG 16 16 36146 1 . LYS 17 17 36146 1 . PHE 18 18 36146 1 . GLY 19 19 36146 1 . ARG 20 20 36146 1 . ARG 21 21 36146 1 stop_ save_ #################### # Natural source # #################### save_natural_source _Entity_natural_src_list.Sf_category natural_source _Entity_natural_src_list.Sf_framecode natural_source _Entity_natural_src_list.Entry_ID 36146 _Entity_natural_src_list.ID 1 loop_ _Entity_natural_src.ID _Entity_natural_src.Entity_ID _Entity_natural_src.Entity_label _Entity_natural_src.Entity_chimera_segment_ID _Entity_natural_src.NCBI_taxonomy_ID _Entity_natural_src.Type _Entity_natural_src.Common _Entity_natural_src.Organism_name_scientific _Entity_natural_src.Organism_name_common _Entity_natural_src.Organism_acronym _Entity_natural_src.ICTVdb_decimal_code _Entity_natural_src.Superkingdom _Entity_natural_src.Kingdom _Entity_natural_src.Genus _Entity_natural_src.Species _Entity_natural_src.Strain _Entity_natural_src.Variant _Entity_natural_src.Organ _Entity_natural_src.Tissue _Entity_natural_src.Tissue_fraction _Entity_natural_src.Cell_line _Entity_natural_src.Cell_type _Entity_natural_src.ATCC_number _Entity_natural_src.Organelle _Entity_natural_src.Secretion _Entity_natural_src.Plasmid _Entity_natural_src.Gene_mnemonic _Entity_natural_src.Details _Entity_natural_src.Entry_ID _Entity_natural_src.Entity_natural_src_list_ID 1 1 $entity_1 . 7739 organism . 'Branchiostoma floridae' 'Florida lancelet' . . Eukaryota Metazoa Branchiostoma floridae . . . . . . . . . . . . . 36146 1 stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Entity_experimental_src_list.Sf_category experimental_source _Entity_experimental_src_list.Sf_framecode experimental_source _Entity_experimental_src_list.Entry_ID 36146 _Entity_experimental_src_list.ID 1 loop_ _Entity_experimental_src.ID _Entity_experimental_src.Entity_ID _Entity_experimental_src.Entity_label _Entity_experimental_src.Entity_chimera_segment_ID _Entity_experimental_src.Production_method _Entity_experimental_src.Host_org_scientific_name _Entity_experimental_src.Host_org_name_common _Entity_experimental_src.Host_org_details _Entity_experimental_src.Host_org_NCBI_taxonomy_ID _Entity_experimental_src.Host_org_genus _Entity_experimental_src.Host_org_species _Entity_experimental_src.Host_org_strain _Entity_experimental_src.Host_org_variant _Entity_experimental_src.Host_org_ATCC_number _Entity_experimental_src.Vector_type _Entity_experimental_src.PDBview_host_org_vector_name _Entity_experimental_src.PDBview_plasmid_name _Entity_experimental_src.Vector_name _Entity_experimental_src.Vector_details _Entity_experimental_src.Vendor_name _Entity_experimental_src.Details _Entity_experimental_src.Entry_ID _Entity_experimental_src.Entity_experimental_src_list_ID 1 1 $entity_1 . 'chemical synthesis' . . . . . . . . . . . . . . . . 36146 1 stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Sample.Sf_category sample _Sample.Sf_framecode sample_1 _Sample.Entry_ID 36146 _Sample.ID 1 _Sample.Type solution _Sample.Sub_type . _Sample.Details '2 mM WT peptides, 10 mM pH 6.0 sodium phosphate, 50 mM pH 6.0 sodium chloride, 10 % pH 6.0 D2O, 90% H2O/10% D2O' _Sample.Aggregate_sample_number . _Sample.Solvent_system '90% H2O/10% D2O' _Sample.Preparation_date . _Sample.Preparation_expiration_date . _Sample.Polycrystallization_protocol . _Sample.Single_crystal_protocol . _Sample.Crystal_grow_apparatus . _Sample.Crystal_grow_atmosphere . _Sample.Crystal_grow_details . _Sample.Crystal_grow_method . _Sample.Crystal_grow_method_cit_ID . _Sample.Crystal_grow_pH . _Sample.Crystal_grow_pH_range . _Sample.Crystal_grow_pressure . _Sample.Crystal_grow_pressure_esd . _Sample.Crystal_grow_seeding . _Sample.Crystal_grow_seeding_cit_ID . _Sample.Crystal_grow_temp . _Sample.Crystal_grow_temp_details . _Sample.Crystal_grow_temp_esd . _Sample.Crystal_grow_time . _Sample.Oriented_sample_prep_protocol . _Sample.Lyophilization_cryo_protectant . _Sample.Storage_protocol . loop_ _Sample_component.ID _Sample_component.Mol_common_name _Sample_component.Isotopic_labeling _Sample_component.Assembly_ID _Sample_component.Assembly_label _Sample_component.Entity_ID _Sample_component.Entity_label _Sample_component.Product_ID _Sample_component.Type _Sample_component.Concentration_val _Sample_component.Concentration_val_min _Sample_component.Concentration_val_max _Sample_component.Concentration_val_units _Sample_component.Concentration_val_err _Sample_component.Vendor _Sample_component.Vendor_product_name _Sample_component.Vendor_product_code _Sample_component.Entry_ID _Sample_component.Sample_ID 1 peptides 'natural abundance' 1 $assembly 1 $entity_1 . protein 2 . . mM . . . . 36146 1 2 'sodium chloride' 'natural abundance' . . . . . salt 50 . . mM . . . . 36146 1 3 'sodium phosphate' 'natural abundance' . . . . . buffer 10 . . mM . . . . 36146 1 4 H2O 'natural abundace' . . . . . solvent 90 . . % . . . . 36146 1 5 D2O [U-2H] . . . . . solvent 10 . . % . . . . 36146 1 stop_ save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Sample_condition_list.Sf_category sample_conditions _Sample_condition_list.Sf_framecode sample_conditions_1 _Sample_condition_list.Entry_ID 36146 _Sample_condition_list.ID 1 _Sample_condition_list.Details . loop_ _Sample_condition_variable.Type _Sample_condition_variable.Val _Sample_condition_variable.Val_err _Sample_condition_variable.Val_units _Sample_condition_variable.Entry_ID _Sample_condition_variable.Sample_condition_list_ID 'ionic strength' 50 . mM 36146 1 pH 6.0 . pH 36146 1 pressure 1 . atm 36146 1 temperature 298.0 . K 36146 1 stop_ save_ ############################ # Computer software used # ############################ save_software_1 _Software.Sf_category software _Software.Sf_framecode software_1 _Software.Entry_ID 36146 _Software.ID 1 _Software.Type . _Software.Name CYANA _Software.Version 2.1 _Software.DOI . _Software.Details . loop_ _Vendor.Name _Vendor.Address _Vendor.Electronic_address _Vendor.Entry_ID _Vendor.Software_ID 'Guntert, Mumenthaler and Wuthrich' . . 36146 1 stop_ loop_ _Task.Task _Task.Entry_ID _Task.Software_ID 'structure calculation' 36146 1 stop_ save_ save_software_2 _Software.Sf_category software _Software.Sf_framecode software_2 _Software.Entry_ID 36146 _Software.ID 2 _Software.Type . _Software.Name Molmol _Software.Version . _Software.DOI . _Software.Details . loop_ _Vendor.Name _Vendor.Address _Vendor.Electronic_address _Vendor.Entry_ID _Vendor.Software_ID 'Koradi, Billeter and Wuthrich' . . 36146 2 stop_ loop_ _Task.Task _Task.Entry_ID _Task.Software_ID 'chemical shift assignment' 36146 2 stop_ save_ save_software_3 _Software.Sf_category software _Software.Sf_framecode software_3 _Software.Entry_ID 36146 _Software.ID 3 _Software.Type . _Software.Name NMRView _Software.Version . _Software.DOI . _Software.Details . loop_ _Vendor.Name _Vendor.Address _Vendor.Electronic_address _Vendor.Entry_ID _Vendor.Software_ID 'Johnson, One Moon Scientific' . . 36146 3 stop_ loop_ _Task.Task _Task.Entry_ID _Task.Software_ID 'peak picking' 36146 3 stop_ save_ save_software_4 _Software.Sf_category software _Software.Sf_framecode software_4 _Software.Entry_ID 36146 _Software.ID 4 _Software.Type . _Software.Name 'X-PLOR NIH' _Software.Version . _Software.DOI . _Software.Details . loop_ _Vendor.Name _Vendor.Address _Vendor.Electronic_address _Vendor.Entry_ID _Vendor.Software_ID 'Schwieters, Kuszewski, Tjandra and Clore' . . 36146 4 stop_ loop_ _Task.Task _Task.Entry_ID _Task.Software_ID refinement 36146 4 stop_ save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _NMR_spectrometer.Sf_category NMR_spectrometer _NMR_spectrometer.Sf_framecode NMR_spectrometer_1 _NMR_spectrometer.Entry_ID 36146 _NMR_spectrometer.ID 1 _NMR_spectrometer.Details . _NMR_spectrometer.Manufacturer Bruker _NMR_spectrometer.Model Avance _NMR_spectrometer.Serial_number . _NMR_spectrometer.Field_strength 600 save_ save_NMR_spectrometer_list _NMR_spectrometer_list.Sf_category NMR_spectrometer_list _NMR_spectrometer_list.Sf_framecode NMR_spectrometer_list _NMR_spectrometer_list.Entry_ID 36146 _NMR_spectrometer_list.ID 1 loop_ _NMR_spectrometer_view.ID _NMR_spectrometer_view.Name _NMR_spectrometer_view.Manufacturer _NMR_spectrometer_view.Model _NMR_spectrometer_view.Serial_number _NMR_spectrometer_view.Field_strength _NMR_spectrometer_view.Details _NMR_spectrometer_view.Citation_ID _NMR_spectrometer_view.Citation_label _NMR_spectrometer_view.Entry_ID _NMR_spectrometer_view.NMR_spectrometer_list_ID 1 NMR_spectrometer_1 Bruker Avance . 600 . . . 36146 1 stop_ save_ ############################# # NMR applied experiments # ############################# save_experiment_list _Experiment_list.Sf_category experiment_list _Experiment_list.Sf_framecode experiment_list _Experiment_list.Entry_ID 36146 _Experiment_list.ID 1 _Experiment_list.Details . loop_ _Experiment.ID _Experiment.Name _Experiment.Raw_data_flag _Experiment.NMR_spec_expt_ID _Experiment.NMR_spec_expt_label _Experiment.MS_expt_ID _Experiment.MS_expt_label _Experiment.SAXS_expt_ID _Experiment.SAXS_expt_label _Experiment.FRET_expt_ID _Experiment.FRET_expt_label _Experiment.EMR_expt_ID _Experiment.EMR_expt_label _Experiment.Sample_ID _Experiment.Sample_label _Experiment.Sample_state _Experiment.Sample_volume _Experiment.Sample_volume_units _Experiment.Sample_condition_list_ID _Experiment.Sample_condition_list_label _Experiment.Sample_spinning_rate _Experiment.Sample_angle _Experiment.NMR_tube_type _Experiment.NMR_spectrometer_ID _Experiment.NMR_spectrometer_label _Experiment.NMR_spectrometer_probe_ID _Experiment.NMR_spectrometer_probe_label _Experiment.NMR_spectral_processing_ID _Experiment.NMR_spectral_processing_label _Experiment.Mass_spectrometer_ID _Experiment.Mass_spectrometer_label _Experiment.Xray_instrument_ID _Experiment.Xray_instrument_label _Experiment.Fluorescence_instrument_ID _Experiment.Fluorescence_instrument_label _Experiment.EMR_instrument_ID _Experiment.EMR_instrument_label _Experiment.Chromatographic_system_ID _Experiment.Chromatographic_system_label _Experiment.Chromatographic_column_ID _Experiment.Chromatographic_column_label _Experiment.Entry_ID _Experiment.Experiment_list_ID 1 '2D 1H-1H TOCSY' no . . . . . . . . . . 1 $sample_1 isotropic . . 1 $sample_conditions_1 . . . 1 $NMR_spectrometer_1 . . . . . . . . . . . . . . . . 36146 1 2 '2D 1H-1H NOESY' no . . . . . . . . . . 1 $sample_1 isotropic . . 1 $sample_conditions_1 . . . 1 $NMR_spectrometer_1 . . . . . . . . . . . . . . . . 36146 1 3 '2D DQF-COSY' no . . . . . . . . . . 1 $sample_1 isotropic . . 1 $sample_conditions_1 . . . 1 $NMR_spectrometer_1 . . . . . . . . . . . . . . . . 36146 1 stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chem_shift_reference_1 _Chem_shift_reference.Sf_category chem_shift_reference _Chem_shift_reference.Sf_framecode chem_shift_reference_1 _Chem_shift_reference.Entry_ID 36146 _Chem_shift_reference.ID 1 _Chem_shift_reference.Details . loop_ _Chem_shift_ref.Atom_type _Chem_shift_ref.Atom_isotope_number _Chem_shift_ref.Mol_common_name _Chem_shift_ref.Atom_group _Chem_shift_ref.Concentration_val _Chem_shift_ref.Concentration_units _Chem_shift_ref.Solvent _Chem_shift_ref.Rank _Chem_shift_ref.Chem_shift_units _Chem_shift_ref.Chem_shift_val _Chem_shift_ref.Ref_method _Chem_shift_ref.Ref_type _Chem_shift_ref.Indirect_shift_ratio _Chem_shift_ref.External_ref_loc _Chem_shift_ref.External_ref_sample_geometry _Chem_shift_ref.External_ref_axis _Chem_shift_ref.Ref_correction_type _Chem_shift_ref.Correction_val _Chem_shift_ref.Entry_ID _Chem_shift_ref.Chem_shift_reference_ID H 1 DSS 'methyl protons' . . . . ppm 0.0 internal direct 1 . . . . . 36146 1 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_chemical_shifts_1 _Assigned_chem_shift_list.Sf_category assigned_chemical_shifts _Assigned_chem_shift_list.Sf_framecode assigned_chemical_shifts_1 _Assigned_chem_shift_list.Entry_ID 36146 _Assigned_chem_shift_list.ID 1 _Assigned_chem_shift_list.Sample_condition_list_ID 1 _Assigned_chem_shift_list.Sample_condition_list_label $sample_conditions_1 _Assigned_chem_shift_list.Chem_shift_reference_ID 1 _Assigned_chem_shift_list.Chem_shift_reference_label $chem_shift_reference_1 _Assigned_chem_shift_list.Chem_shift_1H_err 0.01 _Assigned_chem_shift_list.Chem_shift_13C_err . _Assigned_chem_shift_list.Chem_shift_15N_err . _Assigned_chem_shift_list.Chem_shift_31P_err . _Assigned_chem_shift_list.Chem_shift_2H_err . _Assigned_chem_shift_list.Chem_shift_19F_err . _Assigned_chem_shift_list.Error_derivation_method . _Assigned_chem_shift_list.Details . _Assigned_chem_shift_list.Text_data_format . _Assigned_chem_shift_list.Text_data . loop_ _Chem_shift_experiment.Experiment_ID _Chem_shift_experiment.Experiment_name _Chem_shift_experiment.Sample_ID _Chem_shift_experiment.Sample_label _Chem_shift_experiment.Sample_state _Chem_shift_experiment.Entry_ID _Chem_shift_experiment.Assigned_chem_shift_list_ID 1 '2D 1H-1H TOCSY' 1 $sample_1 isotropic 36146 1 2 '2D 1H-1H NOESY' 1 $sample_1 isotropic 36146 1 3 '2D DQF-COSY' 1 $sample_1 isotropic 36146 1 stop_ loop_ _Atom_chem_shift.ID _Atom_chem_shift.Assembly_atom_ID _Atom_chem_shift.Entity_assembly_ID _Atom_chem_shift.Entity_ID _Atom_chem_shift.Comp_index_ID _Atom_chem_shift.Seq_ID _Atom_chem_shift.Comp_ID _Atom_chem_shift.Atom_ID _Atom_chem_shift.Atom_type _Atom_chem_shift.Atom_isotope_number _Atom_chem_shift.Val _Atom_chem_shift.Val_err _Atom_chem_shift.Assign_fig_of_merit _Atom_chem_shift.Ambiguity_code _Atom_chem_shift.Ambiguity_set_ID _Atom_chem_shift.Occupancy _Atom_chem_shift.Resonance_ID _Atom_chem_shift.Auth_entity_assembly_ID _Atom_chem_shift.Auth_asym_ID _Atom_chem_shift.Auth_seq_ID _Atom_chem_shift.Auth_comp_ID _Atom_chem_shift.Auth_atom_ID _Atom_chem_shift.Details _Atom_chem_shift.Entry_ID _Atom_chem_shift.Assigned_chem_shift_list_ID 1 . 1 1 3 3 CYS H H 1 8.1970 0.01 . 1 . . . . A 3 CYS H . 36146 1 2 . 1 1 3 3 CYS HA H 1 4.6340 0.01 . 1 . . . . A 3 CYS HA . 36146 1 3 . 1 1 3 3 CYS HB2 H 1 3.1760 0.01 . 2 . . . . A 3 CYS HB2 . 36146 1 4 . 1 1 3 3 CYS HB3 H 1 2.5930 0.01 . 2 . . . . A 3 CYS HB3 . 36146 1 5 . 1 1 4 4 ALA H H 1 7.5860 0.01 . 1 . . . . A 4 ALA H . 36146 1 6 . 1 1 4 4 ALA HA H 1 4.1500 0.01 . 1 . . . . A 4 ALA HA . 36146 1 7 . 1 1 4 4 ALA HB1 H 1 1.4200 0.01 . 1 . . . . A 4 ALA HB1 . 36146 1 8 . 1 1 4 4 ALA HB2 H 1 1.4200 0.01 . 1 . . . . A 4 ALA HB2 . 36146 1 9 . 1 1 4 4 ALA HB3 H 1 1.4200 0.01 . 1 . . . . A 4 ALA HB3 . 36146 1 10 . 1 1 5 5 SER H H 1 7.9020 0.01 . 1 . . . . A 5 SER H . 36146 1 11 . 1 1 5 5 SER HA H 1 4.4890 0.01 . 1 . . . . A 5 SER HA . 36146 1 12 . 1 1 5 5 SER HB2 H 1 4.1020 0.01 . 2 . . . . A 5 SER HB2 . 36146 1 13 . 1 1 5 5 SER HB3 H 1 3.9390 0.01 . 2 . . . . A 5 SER HB3 . 36146 1 14 . 1 1 6 6 LEU H H 1 8.4780 0.01 . 1 . . . . A 6 LEU H . 36146 1 15 . 1 1 6 6 LEU HA H 1 4.0560 0.01 . 1 . . . . A 6 LEU HA . 36146 1 16 . 1 1 6 6 LEU HB2 H 1 1.6810 0.01 . 2 . . . . A 6 LEU HB2 . 36146 1 17 . 1 1 6 6 LEU HB3 H 1 1.5870 0.01 . 2 . . . . A 6 LEU HB3 . 36146 1 18 . 1 1 6 6 LEU HG H 1 1.5900 0.01 . 1 . . . . A 6 LEU HG . 36146 1 19 . 1 1 6 6 LEU HD11 H 1 0.8630 0.01 . 2 . . . . A 6 LEU HD11 . 36146 1 20 . 1 1 6 6 LEU HD12 H 1 0.8630 0.01 . 2 . . . . A 6 LEU HD12 . 36146 1 21 . 1 1 6 6 LEU HD13 H 1 0.8630 0.01 . 2 . . . . A 6 LEU HD13 . 36146 1 22 . 1 1 6 6 LEU HD21 H 1 0.9150 0.01 . 2 . . . . A 6 LEU HD21 . 36146 1 23 . 1 1 6 6 LEU HD22 H 1 0.9150 0.01 . 2 . . . . A 6 LEU HD22 . 36146 1 24 . 1 1 6 6 LEU HD23 H 1 0.9150 0.01 . 2 . . . . A 6 LEU HD23 . 36146 1 25 . 1 1 7 7 ARG H H 1 8.3480 0.01 . 1 . . . . A 7 ARG H . 36146 1 26 . 1 1 7 7 ARG HA H 1 4.1500 0.01 . 1 . . . . A 7 ARG HA . 36146 1 27 . 1 1 7 7 ARG HB2 H 1 1.8200 0.01 . 2 . . . . A 7 ARG HB2 . 36146 1 28 . 1 1 7 7 ARG HB3 H 1 1.8200 0.01 . 2 . . . . A 7 ARG HB3 . 36146 1 29 . 1 1 7 7 ARG HG2 H 1 1.6430 0.01 . 2 . . . . A 7 ARG HG2 . 36146 1 30 . 1 1 7 7 ARG HG3 H 1 1.5640 0.01 . 2 . . . . A 7 ARG HG3 . 36146 1 31 . 1 1 7 7 ARG HD2 H 1 3.1780 0.01 . 2 . . . . A 7 ARG HD2 . 36146 1 32 . 1 1 7 7 ARG HD3 H 1 3.1780 0.01 . 2 . . . . A 7 ARG HD3 . 36146 1 33 . 1 1 7 7 ARG HE H 1 7.2860 0.01 . 1 . . . . A 7 ARG HE . 36146 1 34 . 1 1 8 8 ALA H H 1 8.0030 0.01 . 1 . . . . A 8 ALA H . 36146 1 35 . 1 1 8 8 ALA HA H 1 4.1050 0.01 . 1 . . . . A 8 ALA HA . 36146 1 36 . 1 1 8 8 ALA HB1 H 1 1.5300 0.01 . 1 . . . . A 8 ALA HB1 . 36146 1 37 . 1 1 8 8 ALA HB2 H 1 1.5300 0.01 . 1 . . . . A 8 ALA HB2 . 36146 1 38 . 1 1 8 8 ALA HB3 H 1 1.5300 0.01 . 1 . . . . A 8 ALA HB3 . 36146 1 39 . 1 1 9 9 ARG H H 1 7.9910 0.01 . 1 . . . . A 9 ARG H . 36146 1 40 . 1 1 9 9 ARG HA H 1 3.8940 0.01 . 1 . . . . A 9 ARG HA . 36146 1 41 . 1 1 9 9 ARG HB2 H 1 1.8250 0.01 . 2 . . . . A 9 ARG HB2 . 36146 1 42 . 1 1 9 9 ARG HB3 H 1 1.8240 0.01 . 2 . . . . A 9 ARG HB3 . 36146 1 43 . 1 1 9 9 ARG HG2 H 1 1.7180 0.01 . 2 . . . . A 9 ARG HG2 . 36146 1 44 . 1 1 9 9 ARG HG3 H 1 1.5840 0.01 . 2 . . . . A 9 ARG HG3 . 36146 1 45 . 1 1 9 9 ARG HD2 H 1 3.1160 0.01 . 2 . . . . A 9 ARG HD2 . 36146 1 46 . 1 1 9 9 ARG HD3 H 1 3.1160 0.01 . 2 . . . . A 9 ARG HD3 . 36146 1 47 . 1 1 9 9 ARG HE H 1 7.0830 0.01 . 1 . . . . A 9 ARG HE . 36146 1 48 . 1 1 10 10 HIS H H 1 7.9010 0.01 . 1 . . . . A 10 HIS H . 36146 1 49 . 1 1 10 10 HIS HA H 1 4.6950 0.01 . 1 . . . . A 10 HIS HA . 36146 1 50 . 1 1 10 10 HIS HB2 H 1 3.2880 0.01 . 2 . . . . A 10 HIS HB2 . 36146 1 51 . 1 1 10 10 HIS HB3 H 1 3.1780 0.01 . 2 . . . . A 10 HIS HB3 . 36146 1 52 . 1 1 11 11 THR H H 1 7.8260 0.01 . 1 . . . . A 11 THR H . 36146 1 53 . 1 1 11 11 THR HA H 1 4.4410 0.01 . 1 . . . . A 11 THR HA . 36146 1 54 . 1 1 11 11 THR HB H 1 4.1800 0.01 . 1 . . . . A 11 THR HB . 36146 1 55 . 1 1 11 11 THR HG21 H 1 1.1720 0.01 . 1 . . . . A 11 THR HG21 . 36146 1 56 . 1 1 11 11 THR HG22 H 1 1.1720 0.01 . 1 . . . . A 11 THR HG22 . 36146 1 57 . 1 1 11 11 THR HG23 H 1 1.1720 0.01 . 1 . . . . A 11 THR HG23 . 36146 1 58 . 1 1 12 12 ILE H H 1 8.2300 0.01 . 1 . . . . A 12 ILE H . 36146 1 59 . 1 1 12 12 ILE HA H 1 4.5950 0.01 . 1 . . . . A 12 ILE HA . 36146 1 60 . 1 1 12 12 ILE HB H 1 1.9440 0.01 . 1 . . . . A 12 ILE HB . 36146 1 61 . 1 1 12 12 ILE HG12 H 1 1.6120 0.01 . 2 . . . . A 12 ILE HG12 . 36146 1 62 . 1 1 12 12 ILE HG13 H 1 1.1760 0.01 . 2 . . . . A 12 ILE HG13 . 36146 1 63 . 1 1 12 12 ILE HG21 H 1 0.9420 0.01 . 1 . . . . A 12 ILE HG21 . 36146 1 64 . 1 1 12 12 ILE HG22 H 1 0.9420 0.01 . 1 . . . . A 12 ILE HG22 . 36146 1 65 . 1 1 12 12 ILE HG23 H 1 0.9420 0.01 . 1 . . . . A 12 ILE HG23 . 36146 1 66 . 1 1 12 12 ILE HD11 H 1 0.8520 0.01 . 1 . . . . A 12 ILE HD11 . 36146 1 67 . 1 1 12 12 ILE HD12 H 1 0.8520 0.01 . 1 . . . . A 12 ILE HD12 . 36146 1 68 . 1 1 12 12 ILE HD13 H 1 0.8520 0.01 . 1 . . . . A 12 ILE HD13 . 36146 1 69 . 1 1 13 13 PRO HA H 1 4.1200 0.01 . 1 . . . . A 13 PRO HA . 36146 1 70 . 1 1 13 13 PRO HB2 H 1 2.3690 0.01 . 2 . . . . A 13 PRO HB2 . 36146 1 71 . 1 1 13 13 PRO HB3 H 1 2.0000 0.01 . 2 . . . . A 13 PRO HB3 . 36146 1 72 . 1 1 13 13 PRO HG2 H 1 2.1800 0.01 . 2 . . . . A 13 PRO HG2 . 36146 1 73 . 1 1 13 13 PRO HG3 H 1 2.0000 0.01 . 2 . . . . A 13 PRO HG3 . 36146 1 74 . 1 1 13 13 PRO HD2 H 1 3.8700 0.01 . 2 . . . . A 13 PRO HD2 . 36146 1 75 . 1 1 13 13 PRO HD3 H 1 3.8700 0.01 . 2 . . . . A 13 PRO HD3 . 36146 1 76 . 1 1 14 14 GLN H H 1 8.7350 0.01 . 1 . . . . A 14 GLN H . 36146 1 77 . 1 1 14 14 GLN HA H 1 4.2140 0.01 . 1 . . . . A 14 GLN HA . 36146 1 78 . 1 1 14 14 GLN HB2 H 1 2.0730 0.01 . 2 . . . . A 14 GLN HB2 . 36146 1 79 . 1 1 14 14 GLN HB3 H 1 2.0730 0.01 . 2 . . . . A 14 GLN HB3 . 36146 1 80 . 1 1 14 14 GLN HG2 H 1 2.4250 0.01 . 2 . . . . A 14 GLN HG2 . 36146 1 81 . 1 1 14 14 GLN HG3 H 1 2.3740 0.01 . 2 . . . . A 14 GLN HG3 . 36146 1 82 . 1 1 14 14 GLN HE21 H 1 6.8950 0.01 . 2 . . . . A 14 GLN HE21 . 36146 1 83 . 1 1 14 14 GLN HE22 H 1 7.5540 0.01 . 2 . . . . A 14 GLN HE22 . 36146 1 84 . 1 1 15 15 CYS H H 1 8.0740 0.01 . 1 . . . . A 15 CYS H . 36146 1 85 . 1 1 15 15 CYS HA H 1 4.8850 0.01 . 1 . . . . A 15 CYS HA . 36146 1 86 . 1 1 15 15 CYS HB2 H 1 3.1640 0.01 . 2 . . . . A 15 CYS HB2 . 36146 1 87 . 1 1 15 15 CYS HB3 H 1 3.0240 0.01 . 2 . . . . A 15 CYS HB3 . 36146 1 88 . 1 1 16 16 ARG H H 1 7.7250 0.01 . 1 . . . . A 16 ARG H . 36146 1 89 . 1 1 16 16 ARG HA H 1 4.2800 0.01 . 1 . . . . A 16 ARG HA . 36146 1 90 . 1 1 16 16 ARG HB2 H 1 1.8500 0.01 . 2 . . . . A 16 ARG HB2 . 36146 1 91 . 1 1 16 16 ARG HB3 H 1 1.8100 0.01 . 2 . . . . A 16 ARG HB3 . 36146 1 92 . 1 1 16 16 ARG HG2 H 1 1.6570 0.01 . 2 . . . . A 16 ARG HG2 . 36146 1 93 . 1 1 16 16 ARG HG3 H 1 1.6380 0.01 . 2 . . . . A 16 ARG HG3 . 36146 1 94 . 1 1 16 16 ARG HD2 H 1 3.1400 0.01 . 2 . . . . A 16 ARG HD2 . 36146 1 95 . 1 1 16 16 ARG HD3 H 1 3.1400 0.01 . 2 . . . . A 16 ARG HD3 . 36146 1 96 . 1 1 16 16 ARG HE H 1 7.1630 0.01 . 1 . . . . A 16 ARG HE . 36146 1 97 . 1 1 17 17 LYS H H 1 8.0980 0.01 . 1 . . . . A 17 LYS H . 36146 1 98 . 1 1 17 17 LYS HA H 1 4.2230 0.01 . 1 . . . . A 17 LYS HA . 36146 1 99 . 1 1 17 17 LYS HB2 H 1 1.6930 0.01 . 2 . . . . A 17 LYS HB2 . 36146 1 100 . 1 1 17 17 LYS HB3 H 1 1.7000 0.01 . 2 . . . . A 17 LYS HB3 . 36146 1 101 . 1 1 17 17 LYS HG2 H 1 1.2000 0.01 . 2 . . . . A 17 LYS HG2 . 36146 1 102 . 1 1 17 17 LYS HG3 H 1 1.2020 0.01 . 2 . . . . A 17 LYS HG3 . 36146 1 103 . 1 1 17 17 LYS HD2 H 1 1.5830 0.01 . 2 . . . . A 17 LYS HD2 . 36146 1 104 . 1 1 17 17 LYS HD3 H 1 1.5830 0.01 . 2 . . . . A 17 LYS HD3 . 36146 1 105 . 1 1 17 17 LYS HE2 H 1 2.9000 0.01 . 2 . . . . A 17 LYS HE2 . 36146 1 106 . 1 1 17 17 LYS HE3 H 1 2.9000 0.01 . 2 . . . . A 17 LYS HE3 . 36146 1 107 . 1 1 17 17 LYS HZ1 H 1 7.2540 0.01 . 1 . . . . A 17 LYS HZ1 . 36146 1 108 . 1 1 17 17 LYS HZ2 H 1 7.2540 0.01 . 1 . . . . A 17 LYS HZ2 . 36146 1 109 . 1 1 17 17 LYS HZ3 H 1 7.2540 0.01 . 1 . . . . A 17 LYS HZ3 . 36146 1 110 . 1 1 18 18 PHE H H 1 7.9660 0.01 . 1 . . . . A 18 PHE H . 36146 1 111 . 1 1 18 18 PHE HA H 1 4.5530 0.01 . 1 . . . . A 18 PHE HA . 36146 1 112 . 1 1 18 18 PHE HB2 H 1 3.1620 0.01 . 2 . . . . A 18 PHE HB2 . 36146 1 113 . 1 1 18 18 PHE HB3 H 1 3.0340 0.01 . 2 . . . . A 18 PHE HB3 . 36146 1 114 . 1 1 18 18 PHE HD1 H 1 7.2550 0.01 . 3 . . . . A 18 PHE HD1 . 36146 1 115 . 1 1 18 18 PHE HD2 H 1 7.2550 0.01 . 3 . . . . A 18 PHE HD2 . 36146 1 116 . 1 1 18 18 PHE HE1 H 1 7.2550 0.01 . 3 . . . . A 18 PHE HE1 . 36146 1 117 . 1 1 18 18 PHE HE2 H 1 7.2550 0.01 . 3 . . . . A 18 PHE HE2 . 36146 1 118 . 1 1 18 18 PHE HZ H 1 7.3420 0.01 . 1 . . . . A 18 PHE HZ . 36146 1 119 . 1 1 19 19 GLY H H 1 8.2510 0.01 . 1 . . . . A 19 GLY H . 36146 1 120 . 1 1 19 19 GLY HA2 H 1 3.8830 0.01 . 2 . . . . A 19 GLY HA2 . 36146 1 121 . 1 1 19 19 GLY HA3 H 1 3.8830 0.01 . 2 . . . . A 19 GLY HA3 . 36146 1 122 . 1 1 20 20 ARG H H 1 8.1630 0.01 . 1 . . . . A 20 ARG H . 36146 1 123 . 1 1 20 20 ARG HA H 1 4.3060 0.01 . 1 . . . . A 20 ARG HA . 36146 1 124 . 1 1 20 20 ARG HB2 H 1 1.8510 0.01 . 2 . . . . A 20 ARG HB2 . 36146 1 125 . 1 1 20 20 ARG HB3 H 1 1.7670 0.01 . 2 . . . . A 20 ARG HB3 . 36146 1 126 . 1 1 20 20 ARG HG2 H 1 1.6180 0.01 . 2 . . . . A 20 ARG HG2 . 36146 1 127 . 1 1 20 20 ARG HG3 H 1 1.6180 0.01 . 2 . . . . A 20 ARG HG3 . 36146 1 128 . 1 1 20 20 ARG HD2 H 1 3.1740 0.01 . 2 . . . . A 20 ARG HD2 . 36146 1 129 . 1 1 20 20 ARG HD3 H 1 3.1740 0.01 . 2 . . . . A 20 ARG HD3 . 36146 1 130 . 1 1 21 21 ARG H H 1 8.3920 0.01 . 1 . . . . A 21 ARG H . 36146 1 131 . 1 1 21 21 ARG HA H 1 4.2780 0.01 . 1 . . . . A 21 ARG HA . 36146 1 132 . 1 1 21 21 ARG HB2 H 1 1.8540 0.01 . 2 . . . . A 21 ARG HB2 . 36146 1 133 . 1 1 21 21 ARG HB3 H 1 1.7720 0.01 . 2 . . . . A 21 ARG HB3 . 36146 1 134 . 1 1 21 21 ARG HG2 H 1 1.6600 0.01 . 2 . . . . A 21 ARG HG2 . 36146 1 135 . 1 1 21 21 ARG HG3 H 1 1.6120 0.01 . 2 . . . . A 21 ARG HG3 . 36146 1 136 . 1 1 21 21 ARG HD2 H 1 3.1700 0.01 . 2 . . . . A 21 ARG HD2 . 36146 1 137 . 1 1 21 21 ARG HD3 H 1 3.1700 0.01 . 2 . . . . A 21 ARG HD3 . 36146 1 138 . 1 1 21 21 ARG HE H 1 7.2540 0.01 . 1 . . . . A 21 ARG HE . 36146 1 stop_ save_