data_5295 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H Resonance Assignments at 310K and pH 4.93 for E.coli recombinant Brazzein, an Intensely Sweet Protein ; _BMRB_accession_number 5295 _BMRB_flat_file_name bmr5295.str _Entry_type original _Submission_date 2002-01-18 _Accession_date 2002-01-18 _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 Zhao Qin . . 2 Song Jikui . . 3 Markley John L. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 261 "pH NMR parameter values" 6 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2005-09-22 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 5296 'Non-Sweet variant of Brazzein [brazzein-ins(R18a-I18b)]' stop_ _Original_release_date 2002-02-18 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Probing the sweet determinants of brazzein: Wild-type brazzein and a tasteless variant, brazzein-ins(R(18a)-I(18b)), exhibit different pH-dependent NMR chemical shifts ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Zhao Qin . . 2 Song Jikui . . 3 Jin Zheyuan . . 4 Danilova Vicktoria . . 5 Hellekant Goran . . 6 Markley John L. . stop_ _Journal_abbreviation 'Biochem. Biophys. Res. Commun.' _Journal_volume 335 _Journal_issue 1 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 256 _Page_last 263 _Year 2005 _Details . loop_ _Keyword NMR brazzien pKa 'sweet protein' stop_ save_ ################################## # Molecular system description # ################################## save_system_brazzein _Saveframe_category molecular_system _Mol_system_name brazzein _Abbreviation_common brazzein _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label brazzein $brazzein stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all disulfide bound' loop_ _Biological_function 'Elicits intense sweet-tasting response' stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_brazzein _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common brazzein _Name_variant 'sweet protein' _Abbreviation_common bz _Molecular_mass . _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 53 _Mol_residue_sequence ; DKCKKVYENYPVSKCQLANQ CNYDCKLDKHARSGECFYDE 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 HIS 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 2008-08-19 _Sequence_homology_query_revised_last_date 2008-08-19 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 4067 brazzein 100.00 54 100.00 100.00 6.47e-22 BMRB 5723 brazzein 100.00 53 100.00 100.00 6.47e-22 BMRB 5724 brazzein 100.00 53 98.11 98.11 2.69e-21 BMRB 5725 brazzein 100.00 54 100.00 100.00 5.61e-22 BMRB 5726 brazzein 100.00 53 98.11 98.11 2.74e-21 BMRB 5727 brazzein 100.00 53 98.11 98.11 4.75e-21 BMRB 5728 brazzein 100.00 53 98.11 98.11 2.69e-21 PDB 1BRZ 'Solution Structure Of The Sweet Protein Brazzein, Nmr, 43 Structures' 98.11 54 100.00 100.00 2.46e-21 PDB 2BRZ 'Solution Nmr Structure Of The Sweet Protein Brazzein, Minimized Average Structure' 98.11 54 100.00 100.00 2.46e-21 SWISS-PROT P56552 Brazzein 100.00 54 100.00 100.00 5.61e-22 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Organ _Tissue $brazzein 'Joublie fruit' 43545 Eukaryota Viridiplantae Pentadiplandra brazzeana fruit pulp 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 _Isotopic_labeling $brazzein 1 mM . KCl 100 mM . DSS 0.1 mM . stop_ save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_one _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DMX _Field_strength 500 _Details . save_ ############################# # NMR applied experiments # ############################# save_TOCSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name TOCSY _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_conditions_set_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 100 . mM pH 4.93 0.02 pH temperature 310 1 K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chem_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 H 1 'methyl protons' ppm 0.0 internal direct . . . 1.0 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_chemical_shift_assignment_brazzein_ph37310as _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label TOCSY stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $conditions_set_1 _Chem_shift_reference_set_label $chem_shift_reference_1 _Mol_system_component_name 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 H H 8.43 . . 2 . 3 CYS HA H 4.71 . . 3 . 3 CYS HB2 H 3.04 . . 4 . 3 CYS HB3 H 3.32 . . 5 . 4 LYS H H 7.97 . . 6 . 4 LYS HA H 5.55 . . 7 . 4 LYS HB2 H 1.74 . . 8 . 4 LYS HB3 H 1.74 . . 9 . 4 LYS HD2 H 1.50 . . 10 . 4 LYS HD3 H 1.50 . . 11 . 4 LYS HE2 H 2.89 . . 12 . 4 LYS HE3 H 2.89 . . 13 . 4 LYS HG2 H 1.30 . . 14 . 4 LYS HG3 H 1.35 . . 15 . 5 LYS H H 8.96 . . 16 . 5 LYS HA H 4.76 . . 17 . 5 LYS HB2 H 1.81 . . 18 . 5 LYS HB3 H 1.91 . . 19 . 5 LYS HE2 H 2.82 . . 20 . 5 LYS HE3 H 2.82 . . 21 . 5 LYS HG2 H 1.37 . . 22 . 5 LYS HG3 H 1.47 . . 23 . 6 VAL H H 8.78 . . 24 . 6 VAL HA H 3.98 . . 25 . 6 VAL HB H 2.03 . . 26 . 6 VAL HG1 H 1.04 . . 27 . 6 VAL HG2 H 0.90 . . 28 . 7 TYR H H 9.21 . . 29 . 7 TYR HA H 4.43 . . 30 . 7 TYR HB2 H 2.74 . . 31 . 7 TYR HB3 H 3.11 . . 32 . 7 TYR HD1 H 6.79 . . 33 . 7 TYR HD2 H 6.79 . . 34 . 7 TYR HE1 H 6.89 . . 35 . 7 TYR HE2 H 6.89 . . 36 . 8 GLU H H 8.96 . . 37 . 8 GLU HA H 4.19 . . 38 . 8 GLU HB2 H 2.01 . . 39 . 8 GLU HB3 H 2.01 . . 40 . 8 GLU HG2 H 2.40 . . 41 . 8 GLU HG3 H 2.40 . . 42 . 9 ASN H H 9.25 . . 43 . 9 ASN HA H 4.41 . . 44 . 9 ASN HB2 H 3.04 . . 45 . 9 ASN HB3 H 3.10 . . 46 . 9 ASN HD21 H 6.95 . . 47 . 9 ASN HD22 H 7.59 . . 48 . 10 TYR H H 7.51 . . 49 . 10 TYR HA H 4.21 . . 50 . 10 TYR HB2 H 2.77 . . 51 . 10 TYR HB3 H 2.84 . . 52 . 10 TYR HD1 H 7.15 . . 53 . 10 TYR HD2 H 7.15 . . 54 . 10 TYR HE1 H 6.84 . . 55 . 10 TYR HE2 H 6.84 . . 56 . 11 PRO HA H 4.47 . . 57 . 11 PRO HD2 H 3.54 . . 58 . 12 VAL H H 8.07 . . 59 . 12 VAL HA H 3.81 . . 60 . 13 SER H H 8.37 . . 61 . 13 SER HA H 4.15 . . 62 . 13 SER HB2 H 3.84 . . 63 . 13 SER HB3 H 3.84 . . 64 . 14 LYS H H 7.84 . . 65 . 14 LYS HA H 4.06 . . 66 . 14 LYS HB2 H 1.83 . . 67 . 14 LYS HB3 H 1.91 . . 68 . 15 CYS H H 7.86 . . 69 . 15 CYS HA H 4.77 . . 70 . 15 CYS HB2 H 3.02 . . 71 . 15 CYS HB3 H 3.23 . . 72 . 16 GLN H H 7.38 . . 73 . 16 GLN HA H 4.24 . . 74 . 16 GLN HB2 H 2.03 . . 75 . 16 GLN HB3 H 2.28 . . 76 . 16 GLN HE21 H 6.72 . . 77 . 16 GLN HE22 H 7.29 . . 78 . 16 GLN HG3 H 2.41 . . 79 . 17 LEU H H 7.29 . . 80 . 17 LEU HA H 4.53 . . 81 . 17 LEU HB2 H 1.95 . . 82 . 17 LEU HB3 H 1.53 . . 83 . 17 LEU HG H 1.7 . . 84 . 18 ALA H H 8.54 . . 85 . 18 ALA HA H 4.12 . . 86 . 18 ALA HB H 1.39 . . 87 . 19 ASN H H 9.01 . . 88 . 19 ASN HA H 4.67 . . 89 . 19 ASN HB2 H 2.83 . . 90 . 19 ASN HB3 H 2.99 . . 91 . 19 ASN HD21 H 7.04 . . 92 . 19 ASN HD22 H 7.59 . . 93 . 20 GLN H H 7.66 . . 94 . 20 GLN HA H 3.85 . . 95 . 20 GLN HB2 H 2.44 . . 96 . 20 GLN HB3 H 2.05 . . 97 . 20 GLN HE21 H 6.72 . . 98 . 20 GLN HE22 H 7.98 . . 99 . 21 CYS H H 8.48 . . 100 . 21 CYS HA H 4.33 . . 101 . 21 CYS HB2 H 2.54 . . 102 . 21 CYS HB3 H 2.87 . . 103 . 22 ASN H H 7.93 . . 104 . 22 ASN HA H 3.83 . . 105 . 22 ASN HB2 H 2.74 . . 106 . 22 ASN HD21 H 6.73 . . 107 . 22 ASN HD22 H 7.32 . . 108 . 23 TYR H H 8.09 . . 109 . 23 TYR HA H 3.96 . . 110 . 23 TYR HB2 H 3.15 . . 111 . 23 TYR HB3 H 3.20 . . 112 . 23 TYR HD1 H 7.19 . . 113 . 23 TYR HD2 H 7.19 . . 114 . 23 TYR HE1 H 6.85 . . 115 . 23 TYR HE2 H 6.85 . . 116 . 24 ASP H H 9.07 . . 117 . 24 ASP HA H 4.66 . . 118 . 24 ASP HB2 H 2.85 . . 119 . 24 ASP HB3 H 3.16 . . 120 . 25 CYS H H 9.28 . . 121 . 25 CYS HA H 4.15 . . 122 . 25 CYS HB2 H 2.32 . . 123 . 25 CYS HB3 H 2.90 . . 124 . 26 LYS H H 7.66 . . 125 . 26 LYS HA H 4.1 . . 126 . 26 LYS HB2 H 1.43 . . 127 . 26 LYS HB3 H 1.71 . . 128 . 26 LYS HG2 H 1.14 . . 129 . 27 LEU H H 8.22 . . 130 . 27 LEU HA H 4.08 . . 131 . 27 LEU HB2 H 1.65 . . 132 . 28 ASP H H 8.97 . . 133 . 28 ASP HA H 4.53 . . 134 . 28 ASP HB2 H 2.82 . . 135 . 28 ASP HB3 H 3.04 . . 136 . 29 LYS H H 6.63 . . 137 . 29 LYS HA H 4.53 . . 138 . 29 LYS HB2 H 2.26 . . 139 . 29 LYS HB3 H 2.35 . . 140 . 29 LYS HD2 H 1.24 . . 141 . 29 LYS HG2 H 0.87 . . 142 . 30 HIS H H 7.03 . . 143 . 30 HIS HA H 4.60 . . 144 . 30 HIS HB3 H 3.54 . . 145 . 30 HIS HD2 H 7.23 . . 146 . 30 HIS HE1 H 8.61 . . 147 . 31 ALA H H 7.66 . . 148 . 31 ALA HA H 4.10 . . 149 . 31 ALA HB H 0.54 . . 150 . 32 ARG H H 8.22 . . 151 . 32 ARG HA H 4.16 . . 152 . 32 ARG HB2 H 1.49 . . 153 . 32 ARG HB3 H 1.39 . . 154 . 32 ARG HG2 H 1.37 . . 155 . 32 ARG HG3 H 1.48 . . 156 . 33 SER H H 7.99 . . 157 . 33 SER HA H 4.63 . . 158 . 33 SER HB2 H 3.39 . . 159 . 33 SER HB3 H 4.11 . . 160 . 34 GLY H H 8.64 . . 161 . 34 GLY HA2 H 4.86 . . 162 . 34 GLY HA3 H 4.02 . . 163 . 35 GLU H H 8.28 . . 164 . 35 GLU HA H 4.30 . . 165 . 35 GLU HB2 H 1.86 . . 166 . 35 GLU HB3 H 2.05 . . 167 . 36 CYS H H 8.73 . . 168 . 36 CYS HA H 6.15 . . 169 . 36 CYS HB2 H 2.53 . . 170 . 36 CYS HB3 H 3.69 . . 171 . 37 PHE H H 9.03 . . 172 . 37 PHE HA H 4.74 . . 173 . 37 PHE HB2 H 2.75 . . 174 . 37 PHE HB3 H 3.15 . . 175 . 37 PHE HD1 H 7.24 . . 176 . 37 PHE HD2 H 7.24 . . 177 . 37 PHE HE1 H 7.34 . . 178 . 37 PHE HE2 H 7.3 . . 179 . 37 PHE HZ H 7.1 . . 180 . 38 TYR H H 8.60 . . 181 . 38 TYR HA H 4.92 . . 182 . 38 TYR HB2 H 2.86 . . 183 . 38 TYR HB3 H 2.86 . . 184 . 39 ASP H H 8.57 . . 185 . 39 ASP HA H 4.76 . . 186 . 39 ASP HB2 H 3.34 . . 187 . 39 ASP HB3 H 2.78 . . 188 . 40 GLU H H 8.96 . . 189 . 40 GLU HA H 4.18 . . 190 . 41 LYS H H 8.12 . . 191 . 41 LYS HA H 4.41 . . 192 . 41 LYS HB2 H 1.84 . . 193 . 41 LYS HB3 H 2.03 . . 194 . 42 ARG H H 8.30 . . 195 . 42 ARG HA H 3.64 . . 196 . 42 ARG HB2 H 2.05 . . 197 . 42 ARG HB3 H 2.05 . . 198 . 42 ARG HD2 H 3.04 . . 199 . 42 ARG HD3 H 3.04 . . 200 . 42 ARG HE H 6.90 . . 201 . 42 ARG HG3 H 1.50 . . 202 . 43 ASN H H 8.69 . . 203 . 43 ASN HA H 4.96 . . 204 . 43 ASN HB2 H 2.70 . . 205 . 43 ASN HB3 H 2.96 . . 206 . 43 ASN HD21 H 6.95 . . 207 . 43 ASN HD22 H 8.14 . . 208 . 44 LEU H H 8.46 . . 209 . 44 LEU HA H 4.22 . . 210 . 44 LEU HB2 H 1.28 . . 211 . 44 LEU HB3 H 1.51 . . 212 . 44 LEU HD1 H 0.3 . . 213 . 44 LEU HD2 H 0.43 . . 214 . 45 GLN H H 8.47 . . 215 . 45 GLN HA H 4.71 . . 216 . 45 GLN HB2 H 1.88 . . 217 . 45 GLN HB3 H 2.33 . . 218 . 45 GLN HE21 H 7.41 . . 219 . 45 GLN HE22 H 6.69 . . 220 . 45 GLN HG2 H 2.07 . . 221 . 45 GLN HG3 H 2.07 . . 222 . 46 CYS H H 9.19 . . 223 . 46 CYS HA H 4.82 . . 224 . 46 CYS HB2 H 1.69 . . 225 . 46 CYS HB3 H 2.52 . . 226 . 47 ILE H H 8.71 . . 227 . 47 ILE HA H 4.35 . . 228 . 47 ILE HB H 2.16 . . 229 . 47 ILE HD1 H 0.76 . . 230 . 47 ILE HG12 H 1.17 . . 231 . 47 ILE HG13 H 1.6 . . 232 . 48 CYS H H 8.52 . . 233 . 48 CYS HA H 5.30 . . 234 . 48 CYS HB2 H 2.60 . . 235 . 48 CYS HB3 H 2.66 . . 236 . 49 ASP H H 8.83 . . 237 . 49 ASP HA H 5.50 . . 238 . 49 ASP HB2 H 2.36 . . 239 . 49 ASP HB3 H 3.11 . . 240 . 50 TYR H H 9.72 . . 241 . 50 TYR HA H 4.23 . . 242 . 50 TYR HB2 H 2.91 . . 243 . 50 TYR HB3 H 3.46 . . 244 . 50 TYR HD1 H 7.31 . . 245 . 50 TYR HD2 H 7.31 . . 246 . 50 TYR HE1 H 6.71 . . 247 . 50 TYR HE2 H 7.31 . . 248 . 51 CYS H H 9.35 . . 249 . 51 CYS HA H 4.47 . . 250 . 51 CYS HB2 H 3.09 . . 251 . 51 CYS HB3 H 3.53 . . 252 . 52 GLU H H 8.38 . . 253 . 52 GLU HA H 4.28 . . 254 . 52 GLU HB2 H 1.79 . . 255 . 52 GLU HB3 H 2.00 . . 256 . 52 GLU HG2 H 2.19 . . 257 . 52 GLU HG3 H 2.27 . . 258 . 53 TYR H H 7.68 . . 259 . 53 TYR HA H 4.23 . . 260 . 53 TYR HB2 H 2.78 . . 261 . 53 TYR HB3 H 3.13 . . stop_ save_ save_pKa_brazzein_wt _Saveframe_category pKa_value_data_set _Details . loop_ _Experiment_label $TOCSY_1 stop_ loop_ _Sample_label $sample_1 $sample_1 stop_ _Sample_conditions_label $conditions_set_1 _Expt_observed_parameter 'chemical shift' _Text_data_format . _Text_data . save_ save_pKa_chemical_shifts _Saveframe_category pH_NMR_parameter_list _pKa_value_data_set_label $pKa_brazzein_wt _Details . _Text_data_format . _Text_data . save_