data_5296 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H Resonance Assignments at 310K and pH 4.90 for E.coli recombinant Brazzein variant brazzein-ins(R18a-I18b), an Non-Sweet variant of Brazzein ; _BMRB_accession_number 5296 _BMRB_flat_file_name bmr5296.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" 241 "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 5295 'Brazzein; an intensely sweet protein' 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_RI _Saveframe_category molecular_system _Mol_system_name brazzein_RI _Abbreviation_common brazzein_RI _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label brazzein-ins-(R18a-I18b) $brazzein_RI stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all disulfide bound' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_brazzein_RI _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common brazzein-ins(R18a-I18b) _Name_variant 'mutant of sweet protein' _Abbreviation_common bz _Molecular_mass . _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 55 _Mol_residue_sequence ; DKCKKVYENYPVSKCQLRIA NQCNYDCKLDKHARSGECFY DEKRNLQCICDYCEY ; 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 18a ARG 19 18b ILE 20 19 ALA 21 20 ASN 22 21 GLN 23 22 CYS 24 23 ASN 25 24 TYR 26 25 ASP 27 26 CYS 28 27 LYS 29 28 LEU 30 29 ASP 31 30 LYS 32 31 HIS 33 32 ALA 34 33 ARG 35 34 SER 36 35 GLY 37 36 GLU 38 37 CYS 39 38 PHE 40 39 TYR 41 40 ASP 42 41 GLU 43 42 LYS 44 43 ARG 45 44 ASN 46 45 LEU 47 46 GLN 48 47 CYS 49 48 ILE 50 49 CYS 51 50 ASP 52 51 TYR 53 52 CYS 54 53 GLU 55 54 TYR stop_ _Sequence_homology_query_date 2008-08-19 _Sequence_homology_query_revised_last_date 2005-12-09 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 _Details $brazzein_RI 'Joublie fruit' 43545 Eukaryota Viridiplantae Pentadiplandra brazzeana fruit pulp 'Variant of the natural sequence with inserting two residues.' 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_RI '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_RI 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 4.90 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-ins-(R18a-I18b) _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 4 3 CYS H H 8.43 . . 2 4 3 CYS HA H 4.66 . . 3 4 3 CYS HB2 H 3.03 . . 4 4 3 CYS HB3 H 3.31 . . 5 5 4 LYS H H 7.97 . . 6 5 4 LYS HA H 5.54 . . 7 5 4 LYS HB2 H 1.74 . . 8 5 4 LYS HB3 H 1.74 . . 9 5 4 LYS HD2 H 1.51 . . 10 5 4 LYS HD3 H 1.51 . . 11 5 4 LYS HE2 H 2.81 . . 12 5 4 LYS HE3 H 2.81 . . 13 5 4 LYS HG2 H 1.31 . . 14 6 5 LYS H H 8.94 . . 15 6 5 LYS HA H 4.75 . . 16 6 5 LYS HB2 H 1.80 . . 17 6 5 LYS HB3 H 1.91 . . 18 6 5 LYS HE2 H 2.82 . . 19 6 5 LYS HE3 H 2.82 . . 20 6 5 LYS HG2 H 1.35 . . 21 6 5 LYS HG3 H 1.46 . . 22 7 6 VAL H H 8.78 . . 23 7 6 VAL HA H 3.99 . . 24 7 6 VAL HB H 2.03 . . 25 7 6 VAL HG1 H 1.03 . . 26 7 6 VAL HG2 H 0.90 . . 27 8 7 TYR H H 9.19 . . 28 8 7 TYR HA H 4.42 . . 29 8 7 TYR HB2 H 2.76 . . 30 8 7 TYR HB3 H 3.09 . . 31 9 8 GLU H H 8.98 . . 32 9 8 GLU HA H 4.15 . . 33 9 8 GLU HB2 H 2.01 . . 34 9 8 GLU HB3 H 2.01 . . 35 9 8 GLU HG2 H 2.41 . . 36 9 8 GLU HG3 H 2.41 . . 37 10 9 ASN H H 9.25 . . 38 10 9 ASN HA H 4.40 . . 39 10 9 ASN HB2 H 3.01 . . 40 10 9 ASN HB3 H 3.09 . . 41 10 9 ASN HD21 H 6.93 . . 42 10 9 ASN HD22 H 7.59 . . 43 11 10 TYR H H 7.28 . . 44 11 10 TYR HA H 3.91 . . 45 11 10 TYR HB2 H 3.07 . . 46 11 10 TYR HB3 H 3.14 . . 47 13 12 VAL H H 8.10 . . 48 13 12 VAL HA H 3.79 . . 49 14 13 SER H H 8.34 . . 50 14 13 SER HA H 4.15 . . 51 14 13 SER HB2 H 3.84 . . 52 14 13 SER HB3 H 3.84 . . 53 15 14 LYS H H 7.74 . . 54 15 14 LYS HA H 4.15 . . 55 15 14 LYS HB2 H 1.89 . . 56 15 14 LYS HB3 H 1.89 . . 57 15 14 LYS HD2 H 1.26 . . 58 15 14 LYS HD3 H 1.38 . . 59 16 15 CYS H H 7.84 . . 60 16 15 CYS HA H 4.84 . . 61 16 15 CYS HB2 H 3.08 . . 62 17 16 GLN H H 7.51 . . 63 17 16 GLN HA H 4.23 . . 64 17 16 GLN HB2 H 2.05 . . 65 17 16 GLN HB3 H 2.32 . . 66 17 16 GLN HG3 H 2.40 . . 67 18 17 LEU H H 7.32 . . 68 18 17 LEU HA H 4.40 . . 69 18 17 LEU HB2 H 1.82 . . 70 18 17 LEU HG H 1.72 . . 71 18a 18 ARG H H 8.40 . . 72 18a 18 ARG HA H 4.25 . . 73 18a 18 ARG HB2 H 1.87 . . 74 18a 18 ARG HB3 H 1.97 . . 75 18a 18 ARG HG2 H 1.63 . . 76 18a 18 ARG HG3 H 1.73 . . 77 18b 19 ILE H H 7.71 . . 78 18b 19 ILE HA H 4.09 . . 79 18b 19 ILE HD1 H 0.95 . . 80 19 20 ALA H H 7.68 . . 81 19 20 ALA HA H 4.33 . . 82 19 20 ALA HB H 1.38 . . 83 20 21 ASN H H 8.25 . . 84 20 21 ASN HA H 4.75 . . 85 20 21 ASN HB2 H 2.80 . . 86 20 21 ASN HB3 H 3.09 . . 87 20 21 ASN HD21 H 6.85 . . 88 20 21 ASN HD22 H 7.70 . . 89 21 22 GLN H H 7.45 . . 90 21 22 GLN HA H 3.82 . . 91 21 22 GLN HE21 H 6.64 . . 92 21 22 GLN HE22 H 7.7 . . 93 21 22 GLN HG2 H 2.69 . . 94 22 23 CYS H H 8.30 . . 95 22 23 CYS HA H 4.24 . . 96 22 23 CYS HB2 H 2.52 . . 97 22 23 CYS HB3 H 2.85 . . 98 23 24 ASN H H 8.30 . . 99 23 24 ASN HA H 4.24 . . 100 23 24 ASN HB2 H 2.52 . . 101 23 24 ASN HB3 H 2.85 . . 102 24 25 TYR H H 8.04 . . 103 24 25 TYR HA H 4.10 . . 104 24 25 TYR HB3 H 3.19 . . 105 26 27 CYS H H 9.14 . . 106 26 27 CYS HA H 4.13 . . 107 26 27 CYS HB3 H 2.84 . . 108 27 28 LYS H H 7.50 . . 109 27 28 LYS HA H 4.07 . . 110 27 28 LYS HB3 H 1.69 . . 111 28 29 LEU H H 8.20 . . 112 28 29 LEU HA H 4.06 . . 113 28 29 LEU HB2 H 1.64 . . 114 29 30 ASP H H 8.97 . . 115 29 30 ASP HA H 4.53 . . 116 29 30 ASP HB2 H 2.82 . . 117 29 30 ASP HB3 H 3.02 . . 118 30 31 LYS H H 6.60 . . 119 30 31 LYS HA H 4.52 . . 120 30 31 LYS HB2 H 2.25 . . 121 30 31 LYS HB3 H 2.35 . . 122 30 31 LYS HD2 H 1.24 . . 123 30 31 LYS HG2 H 0.86 . . 124 31 32 HIS H H 7.04 . . 125 31 32 HIS HA H 4.58 . . 126 31 32 HIS HB3 H 3.54 . . 127 31 32 HIS HD2 H 7.21 . . 128 31 32 HIS HE1 H 8.60 . . 129 32 33 ALA H H 7.67 . . 130 32 33 ALA HA H 4.09 . . 131 32 33 ALA HB H 0.52 . . 132 33 34 ARG H H 8.23 . . 133 33 34 ARG HA H 4.15 . . 134 33 34 ARG HB2 H 1.47 . . 135 33 34 ARG HB3 H 1.38 . . 136 34 35 SER H H 7.98 . . 137 34 35 SER HA H 4.59 . . 138 34 35 SER HB2 H 3.39 . . 139 34 35 SER HB3 H 4.12 . . 140 35 36 GLY H H 8.65 . . 141 35 36 GLY HA2 H 4.84 . . 142 35 36 GLY HA3 H 3.97 . . 143 36 37 GLU H H 8.32 . . 144 36 37 GLU HA H 4.35 . . 145 36 37 GLU HB2 H 1.88 . . 146 36 37 GLU HB3 H 2.04 . . 147 36 37 GLU HG2 H 2.05 . . 148 36 37 GLU HG3 H 2.05 . . 149 37 38 CYS H H 8.77 . . 150 37 38 CYS HA H 5.96 . . 151 37 38 CYS HB2 H 2.53 . . 152 37 38 CYS HB3 H 3.48 . . 153 38 39 PHE H H 9.16 . . 154 38 39 PHE HA H 4.73 . . 155 38 39 PHE HB2 H 2.75 . . 156 38 39 PHE HB3 H 3.08 . . 157 39 40 TYR H H 8.48 . . 158 39 40 TYR HA H 4.91 . . 159 39 40 TYR HB2 H 2.82 . . 160 40 41 ASP H H 8.52 . . 161 40 41 ASP HA H 4.72 . . 162 40 41 ASP HB2 H 3.32 . . 163 40 41 ASP HB3 H 2.74 . . 164 41 42 GLU H H 8.96 . . 165 41 42 GLU HA H 4.16 . . 166 41 42 GLU HB2 H 2.12 . . 167 41 42 GLU HB3 H 2.39 . . 168 41 42 GLU HG2 H 2.40 . . 169 42 43 LYS H H 8.11 . . 170 42 43 LYS HA H 4.40 . . 171 42 43 LYS HB2 H 1.80 . . 172 42 43 LYS HB3 H 2.04 . . 173 42 43 LYS HD2 H 1.68 . . 174 43 44 ARG H H 8.29 . . 175 43 44 ARG HA H 3.62 . . 176 43 44 ARG HB2 H 2.04 . . 177 43 44 ARG HB3 H 2.04 . . 178 43 44 ARG HD2 H 3.06 . . 179 43 44 ARG HE H 6.91 . . 180 43 44 ARG HG2 H 1.45 . . 181 43 44 ARG HG3 H 1.50 . . 182 44 45 ASN H H 8.68 . . 183 44 45 ASN HA H 4.92 . . 184 44 45 ASN HB2 H 2.69 . . 185 44 45 ASN HB3 H 2.95 . . 186 44 45 ASN HD21 H 6.93 . . 187 44 45 ASN HD22 H 8.14 . . 188 45 46 LEU H H 8.43 . . 189 45 46 LEU HA H 4.25 . . 190 45 46 LEU HB2 H 1.29 . . 191 45 46 LEU HB3 H 1.44 . . 192 45 46 LEU HD1 H 0.33 . . 193 46 47 GLN H H 8.54 . . 194 46 47 GLN HA H 4.68 . . 195 46 47 GLN HB2 H 1.86 . . 196 46 47 GLN HB3 H 2.30 . . 197 46 47 GLN HE21 H 7.38 . . 198 46 47 GLN HE22 H 6.67 . . 199 47 48 CYS H H 9.20 . . 200 47 48 CYS HA H 4.82 . . 201 47 48 CYS HB2 H 1.71 . . 202 47 48 CYS HB3 H 2.47 . . 203 48 49 ILE H H 8.73 . . 204 48 49 ILE HA H 4.35 . . 205 48 49 ILE HB H 2.11 . . 206 48 49 ILE HD1 H 0.74 . . 207 48 49 ILE HG12 H 1.15 . . 208 48 49 ILE HG13 H 1.54 . . 209 49 50 CYS H H 8.52 . . 210 49 50 CYS HA H 5.28 . . 211 49 50 CYS HB2 H 2.58 . . 212 49 50 CYS HB3 H 2.65 . . 213 50 51 ASP H H 8.79 . . 214 50 51 ASP HA H 5.45 . . 215 50 51 ASP HB2 H 2.33 . . 216 50 51 ASP HB3 H 3.11 . . 217 51 52 TYR H H 9.73 . . 218 51 52 TYR HA H 4.22 . . 219 51 52 TYR HB2 H 2.93 . . 220 51 52 TYR HB3 H 3.44 . . 221 51 52 TYR HD2 H 7.32 . . 222 51 52 TYR HE1 H 6.72 . . 223 51 52 TYR HE2 H 6.72 . . 224 52 53 CYS H H 9.36 . . 225 52 53 CYS HA H 4.46 . . 226 52 53 CYS HB2 H 3.09 . . 227 52 53 CYS HB3 H 3.51 . . 228 53 54 GLU H H 8.38 . . 229 53 54 GLU HA H 4.27 . . 230 53 54 GLU HB2 H 1.78 . . 231 53 54 GLU HB3 H 1.99 . . 232 53 54 GLU HG2 H 2.19 . . 233 53 54 GLU HG3 H 2.27 . . 234 54 55 TYR H H 7.68 . . 235 54 55 TYR HA H 4.20 . . 236 54 55 TYR HB2 H 2.78 . . 237 54 55 TYR HB3 H 3.13 . . 238 54 55 TYR HD1 H 7.68 . . 239 54 55 TYR HD2 H 4.20 . . 240 54 55 TYR HE1 H 2.78 . . 241 54 55 TYR HE2 H 3.13 . . stop_ save_ save_pKa_brazzein_RI _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_RI _Details . _Text_data_format . _Text_data . save_