data_5727 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Backbone 1H, 13C, and 15N Chemical Shift Assignments for H31A Brazzein ; _BMRB_accession_number 5727 _BMRB_flat_file_name bmr5727.str _Entry_type original _Submission_date 2003-03-10 _Accession_date 2003-03-10 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details . loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Assadi-Porter Fariba M. . 2 Abilgaard Frits . . 3 Blad Heike . . 4 Markley John L. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 coupling_constants 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 50 "13C chemical shifts" 49 "15N chemical shifts" 50 "coupling constants" 17 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2003-09-15 original author . stop_ loop_ _Related_BMRB_accession_number _Relationship 5723 'wt brazzein' 5724 'R43A brazzein' 5725 'Ala2ins brazzein' 5726 'D50A brazzein' 5728 'R33A brazzein' stop_ save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Correlation of the Sweetness of Variants of the Protein Brazzein with Patterns of Hydrogen Bonds Detected by NMR Spectroscopy ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code 22791616 _PubMed_ID 12732626 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Assadi-Porter Fariba M. . 2 Abilgaard Frits . . 3 Blad Heike . . 4 Markley John L. . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_volume 278 _Journal_issue 33 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 31331 _Page_last 31339 _Year 2003 _Details . loop_ _Keyword taste 'sweet protein' 'hydrogen bond' J-coupling dynamics flexibility rigidity 'proteinase inhibitor' stop_ save_ ################################## # Molecular system description # ################################## save_system_brazzein _Saveframe_category molecular_system _Mol_system_name His31Ala-brazzein _Abbreviation_common brazzein _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label His31Ala-brazzein $brazzein stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'not reported' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_brazzein _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common brazzein _Abbreviation_common bz _Molecular_mass . _Mol_thiol_state 'not reported' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 53 _Mol_residue_sequence ; DKCKKVYENYPVSKCQLANQ CNYDCKLDKAARSGECFYDE KRNLQCICDYCEY ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 2 ASP 2 3 LYS 3 4 CYS 4 5 LYS 5 6 LYS 6 7 VAL 7 8 TYR 8 9 GLU 9 10 ASN 10 11 TYR 11 12 PRO 12 13 VAL 13 14 SER 14 15 LYS 15 16 CYS 16 17 GLN 17 18 LEU 18 19 ALA 19 20 ASN 20 21 GLN 21 22 CYS 22 23 ASN 23 24 TYR 24 25 ASP 25 26 CYS 26 27 LYS 27 28 LEU 28 29 ASP 29 30 LYS 30 31 ALA 31 32 ALA 32 33 ARG 33 34 SER 34 35 GLY 35 36 GLU 36 37 CYS 37 38 PHE 38 39 TYR 39 40 ASP 40 41 GLU 41 42 LYS 42 43 ARG 43 44 ASN 44 45 LEU 45 46 GLN 46 47 CYS 47 48 ILE 48 49 CYS 49 50 ASP 50 51 TYR 51 52 CYS 52 53 GLU 53 54 TYR stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2014-11-09 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value BMRB 16215 wt-brazzein 100.00 53 98.11 98.11 2.90e-27 BMRB 18710 entity 100.00 53 98.11 98.11 2.90e-27 BMRB 4067 brazzein 100.00 54 98.11 98.11 2.96e-27 BMRB 5295 brazzein 100.00 53 98.11 98.11 2.90e-27 BMRB 5723 brazzein 100.00 53 98.11 98.11 2.90e-27 BMRB 5725 brazzein 100.00 54 98.11 98.11 2.63e-27 PDB 1BRZ "Solution Structure Of The Sweet Protein Brazzein, Nmr, 43 Structures" 98.11 54 98.08 98.08 1.70e-26 PDB 2BRZ "Solution Nmr Structure Of The Sweet Protein Brazzein, Minimized Average Structure" 98.11 54 98.08 98.08 1.70e-26 PDB 2KGQ "Refined Solution Structure Of Des-pyro Glu Brazzein" 98.11 53 98.08 98.08 1.66e-26 PDB 2LY5 "Refined Solution Structure Of Recombinant Brazzein" 98.11 53 98.08 98.08 1.66e-26 PDB 2LY6 "Refined Solution Structure Of Recombinant Brazzein At Low Temperature" 98.11 53 98.08 98.08 1.66e-26 PDB 4HE7 "Crystal Structure Of Brazzein" 100.00 54 98.11 98.11 2.96e-27 GB ACK76425 "brazzein [synthetic construct]" 100.00 54 98.11 98.11 2.72e-27 GB AGM38242 "brazzein [synthetic construct]" 100.00 54 98.11 98.11 2.36e-27 SP P56552 "RecName: Full=Defensin-like protein; AltName: Full=Brazzein [Pentadiplandra brazzeana]" 100.00 54 98.11 98.11 2.72e-27 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $brazzein 'Escherichia coli' 562 Eubacteria . Escherichia coli stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Vector_name $brazzein 'recombinant technology' . . . . . stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $brazzein . mM 2.0 4.0 '[U-100% 13C; U-100% 15N]' 2,2-dimethyl-2-silapentane-5-sulfonate 0.1 mM . . . 'sodium azide' 1 mM . . . 'succinic acid' 2 mM . . [U-2H] stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $brazzein 2.0 mM '[U-100% 15N]' stop_ save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DMX _Field_strength 600 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DMX _Field_strength 750 _Details . save_ ############################# # NMR applied experiments # ############################# save_15N-HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name 15N-HSQC _Sample_label . save_ save_HNCO_2 _Saveframe_category NMR_applied_experiment _Experiment_name HNCO _Sample_label . save_ save_HNCOCA_3 _Saveframe_category NMR_applied_experiment _Experiment_name HNCOCA _Sample_label . save_ save_HNCA_4 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA _Sample_label . save_ save_1H-1H-15N_TOCSY_5 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H-15N TOCSY' _Sample_label . save_ save_1H-1H-15N_NOESY_6 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H-15N NOESY' _Sample_label . save_ save_long_range_HNCO_7 _Saveframe_category NMR_applied_experiment _Experiment_name 'long range HNCO' _Sample_label . save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name 15N-HSQC _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name HNCO _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name HNCOCA _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_4 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_5 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H-15N TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_6 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H-15N NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_7 _Saveframe_category NMR_applied_experiment _Experiment_name 'long range HNCO' _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_condition_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 5.2 0.2 n/a temperature 310 1 K 'ionic strength' 2 . mM stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference _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 H 1 'methyl protons' ppm 0.0 internal direct . . . 1.0 DSS N 15 'methyl protons' ppm 0.0 . indirect . . . 0.101329118 DSS C 13 'methyl protons' ppm 0.0 . indirect . . . 0.251449530 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_bz_cs_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $condition_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name His31Ala-brazzein _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 . 3 CYS N N 117.0 . 1 2 . 3 CYS H H 8.45 . 1 3 . 3 CYS C C 174.4 . 1 4 . 4 LYS N N 118.7 . 1 5 . 4 LYS H H 8.0 . 1 6 . 4 LYS C C 176.5 . 1 7 . 5 LYS N N 121.4 . 1 8 . 5 LYS H H 9.01 . 1 9 . 5 LYS C C 176.8 . 1 10 . 6 VAL N N 127.4 . 1 11 . 6 VAL H H 8.74 . 1 12 . 6 VAL C C 175.2 . 1 13 . 7 TYR N N 130.2 . 1 14 . 7 TYR H H 9.22 . 1 15 . 7 TYR C C 175.1 . 1 16 . 8 GLU N N 130.8 . 1 17 . 8 GLU H H 8.85 . 1 18 . 8 GLU C C 177.0 . 1 19 . 9 ASN N N 116.3 . 1 20 . 9 ASN H H 9.25 . 1 21 . 9 ASN C C 174.5 . 1 22 . 10 TYR N N 122.6 . 1 23 . 10 TYR H H 7.51 . 1 24 . 11 PRO C C 177.0 . 1 25 . 12 VAL N N 124.0 . 1 26 . 12 VAL H H 8.05 . 1 27 . 12 VAL C C 178.6 . 1 28 . 13 SER N N 117.5 . 1 29 . 13 SER H H 8.37 . 1 30 . 13 SER C C 177.8 . 1 31 . 14 LYS N N 121.4 . 1 32 . 14 LYS H H 7.87 . 1 33 . 14 LYS C C 178.9 . 1 34 . 15 CYS N N 115.8 . 1 35 . 15 CYS H H 7.85 . 1 36 . 15 CYS C C 175.9 . 1 37 . 16 GLN N N 115.6 . 1 38 . 16 GLN H H 7.36 . 1 39 . 16 GLN C C 175.4 . 1 40 . 17 LEU N N 121.3 . 1 41 . 17 LEU H H 7.30 . 1 42 . 17 LEU C C 176.4 . 1 43 . 18 ALA N N 127.9 . 1 44 . 18 ALA H H 8.52 . 1 45 . 18 ALA C C 179.0 . 1 46 . 19 ASN N N 116.2 . 1 47 . 19 ASN H H 9.06 . 1 48 . 19 ASN C C 175.1 . 1 49 . 20 GLN N N 123.1 . 1 50 . 20 GLN H H 7.67 . 1 51 . 20 GLN C C 176.5 . 1 52 . 21 CYS N N 119.3 . 1 53 . 21 CYS H H 8.43 . 1 54 . 21 CYS C C 176.1 . 1 55 . 22 ASN N N 118.9 . 1 56 . 22 ASN H H 7.91 . 1 57 . 22 ASN C C 175.8 . 1 58 . 23 TYR N N 118.1 . 1 59 . 23 TYR H H 8.11 . 1 60 . 23 TYR C C 177.6 . 1 61 . 24 ASP N N 119.9 . 1 62 . 24 ASP H H 9.06 . 1 63 . 24 ASP C C 179.8 . 1 64 . 25 CYS N N 120.3 . 1 65 . 25 CYS H H 9.24 . 1 66 . 25 CYS C C 177.8 . 1 67 . 26 LYS N N 119.5 . 1 68 . 26 LYS H H 7.64 . 1 69 . 26 LYS C C 179.0 . 1 70 . 27 LEU N N 119.4 . 1 71 . 27 LEU H H 8.23 . 1 72 . 27 LEU C C 178.7 . 1 73 . 28 ASP N N 117.7 . 1 74 . 28 ASP H H 9.10 . 1 75 . 28 ASP C C 178.6 . 1 76 . 29 LYS N N 113.4 . 1 77 . 29 LYS H H 6.80 . 1 78 . 29 LYS C C 176.5 . 1 79 . 30 ALA N N 120.1 . 1 80 . 30 ALA H H 7.40 . 1 81 . 30 ALA C C 175.3 . 1 82 . 31 ALA N N 119.0 . 1 83 . 31 ALA H H 7.60 . 1 84 . 31 ALA C C 175.08 . 1 85 . 32 ARG N N 116.6 . 1 86 . 32 ARG H H 8.20 . 1 87 . 32 ARG C C 177.3 . 1 88 . 33 SER N N 109.4 . 1 89 . 33 SER H H 7.99 . 1 90 . 33 SER C C 173.1 . 1 91 . 34 GLY N N 107.6 . 1 92 . 34 GLY H H 8.60 . 1 93 . 34 GLY C C 169.0 . 1 94 . 35 GLU N N 119.0 . 1 95 . 35 GLU H H 8.27 . 1 96 . 35 GLU C C 174.3 . 1 97 . 36 CYS N N 120.6 . 1 98 . 36 CYS H H 8.75 . 1 99 . 36 CYS C C 174.4 . 1 100 . 37 PHE N N 121.1 . 1 101 . 37 PHE H H 9.04 . 1 102 . 37 PHE C C 175.1 . 1 103 . 38 TYR N N 121.7 . 1 104 . 38 TYR H H 8.62 . 1 105 . 38 TYR C C 176.9 . 1 106 . 39 ASP N N 123.3 . 1 107 . 39 ASP H H 8.59 . 1 108 . 39 ASP C C 178.9 . 1 109 . 40 GLU N N 118.1 . 1 110 . 40 GLU H H 9.05 . 1 111 . 40 GLU C C 177.5 . 1 112 . 41 LYS N N 120.2 . 1 113 . 41 LYS H H 8.14 . 1 114 . 41 LYS C C 175.4 . 1 115 . 42 ARG N N 113.6 . 1 116 . 42 ARG H H 8.30 . 1 117 . 42 ARG C C 175.2 . 1 118 . 43 ASN N N 118.3 . 1 119 . 43 ASN H H 8.73 . 1 120 . 43 ASN C C 175.2 . 1 121 . 44 LEU N N 126.1 . 1 122 . 44 LEU H H 8.45 . 1 123 . 44 LEU C C 175.8 . 1 124 . 45 GLN N N 120.7 . 1 125 . 45 GLN H H 8.46 . 1 126 . 45 GLN C C 174.7 . 1 127 . 46 CYS N N 122.2 . 1 128 . 46 CYS H H 9.17 . 1 129 . 46 CYS C C 172.1 . 1 130 . 47 ILE N N 129.4 . 1 131 . 47 ILE H H 8.73 . 1 132 . 47 ILE C C 175.7 . 1 133 . 48 CYS N N 126.5 . 1 134 . 48 CYS H H 8.60 . 1 135 . 48 CYS C C 173.1 . 1 136 . 49 ASP N N 122.8 . 1 137 . 49 ASP H H 8.83 . 1 138 . 49 ASP C C 176.4 . 1 139 . 50 TYR N N 125.2 . 1 140 . 50 TYR H H 9.64 . 1 141 . 50 TYR C C 175.6 . 1 142 . 51 CYS N N 120.6 . 1 143 . 51 CYS H H 9.56 . 1 144 . 51 CYS C C 173.0 . 1 145 . 52 GLU N N 121.3 . 1 146 . 52 GLU H H 8.35 . 1 147 . 52 GLU C C 174.6 . 1 148 . 53 TYR N N 125.6 . 1 149 . 53 TYR H H 7.65 . 1 stop_ save_ ######################## # Coupling constants # ######################## save_coupling_constants _Saveframe_category coupling_constants _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $condition_1 _Spectrometer_frequency_1H 750 _Mol_system_component_name His31Ala-brazzein _Text_data_format . _Text_data . loop_ _Coupling_constant_ID _Coupling_constant_code _Atom_one_residue_seq_code _Atom_one_residue_label _Atom_one_name _Atom_two_residue_seq_code _Atom_two_residue_label _Atom_two_name _Coupling_constant_value _Coupling_constant_min_value _Coupling_constant_max_value _Coupling_constant_value_error 1 3HJNiCj 5 LYS N 48 CYS C 2.89 . . 0.04 2 3HJNiCj 7 TYR N 46 CYS C 2.83 . . 0.03 3 3HJNiCj 23 TYR N 19 ASN C 2.98 . . 0.05 4 3HJNiCj 24 ASP N 20 GLN C 2.89 . . 0.04 5 3HJNiCj 25 CYS N 21 CYS C 2.81 . . 0.03 6 3HJNiCj 26 LYS N 22 ASN C 3.00 . . 0.07 7 3HJNiCj 27 LEU N 23 TYR C 2.98 . . 0.05 8 3HJNiCj 28 ASP N 24 ASP C 2.88 . . 0.04 9 3HJNiCj 32 ARG N 49 ASP C 3.00 . . 0.06 10 3HJNiCj 35 GLU N 47 ILE C 3.06 . . 0.09 11 3HJNiCj 37 PHE N 45 GLN C 2.89 . . 0.05 12 3HJNiCj 39 ASP N 43 ASN C 2.91 . . 0.05 13 3HJNiCj 45 GLN N 37 PHE C 2.91 . . 0.04 14 3HJNiCj 47 ILE N 35 GLU C 2.94 . . 0.05 15 3HJNiCj 48 CYS N 5 LYS C 2.92 . . 0.05 16 3HJNiCj 49 ASP N 33 SER C 2.92 . . 0.05 17 3HJNiCj 50 TYR N 3 CYS C 2.98 . . 0.04 stop_ save_