data_27493 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H, and 15N Chemical Shift Titration Study of Copper Binding Lipoprotein (bsCopL) ; _BMRB_accession_number 27493 _BMRB_flat_file_name bmr27493.str _Entry_type original _Submission_date 2018-05-23 _Accession_date 2018-05-23 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details ; This is a titration study of bsCopL (residues 83-205) deposited in the Biological Magnetic Resonance Bank (BMRB ID 16942). The titration study was conducted using different concentration of copper sulfate to evaluate the binding properties of copper to bsCopL. The addition of copper results in specific chemical shift perturbation and broadened signals. Additionally, titration experiments reveal that bsCopL has multiple copper binding sites which exhibit multiple copper binding patterns such as weak fast-exchange binding interactions, tight slow-exchange interactions or a combination of fast and slow exchange. ; loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Daigham Nourhan S. . 2 Rosario-Cruz Zuelay . . 3 Eletsky Alexander . . 4 Swapna G.V.T. . . 5 Szyperski Thomas . . 6 Boyd Jeffrey M. . 7 Montelione Gaetano T. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 2 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 130 "15N chemical shifts" 130 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2019-03-26 update BMRB 'update entry citation' 2018-07-06 original author 'original release' stop_ loop_ _Related_BMRB_accession_number _Relationship 16942 'Solution NMR Structure () from B.subtilis, Northeast Structural Genomics Consortium Target Target SR518' stop_ _Original_release_date 2018-05-23 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; The copBL operon protects Staphylococcus aureus from copper toxicity: CopL is an extracellular membrane-associated copper-binding protein ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 30655293 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Rosario-Cruz Zuelay . . 2 Eletsky Alexander . . 3 Daigham Nourhan S. . 4 Al-Tameemi Hassan . . 5 Swapna G . . 6 Kahn Peter C. . 7 Szyperski Thomas . . 8 Montelione Gaetano T. . 9 Boyd Jeffrey M. . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'The Journal of biological chemistry' _Journal_volume 294 _Journal_issue 11 _Journal_ISSN 1083-351X _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 4027 _Page_last 4044 _Year 2019 _Details . loop_ _Keyword Copper Intoxication 'Staphylococcus aureus' Titration bsCopL stop_ save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name bsCopL _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label bsCopL $bsCopL 'copper ion' $entity_CU stop_ _System_molecular_weight 13745.39 _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_bsCopL _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common bsCopL _Molecular_mass 13745.39 _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 126 _Mol_residue_sequence ; SHMKVGSQVIINTSHMKGMK GAEATVTGAYDTTAYVVSYT PTNGGQRVDHHKWVIQEEIK DAGDKTLQPGDQVILEASHM KGMKGATAEIDSAEKTTVYM VDYTSTTSGEKVKNHKWVTE DELSAK ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 80 SER 2 81 HIS 3 82 MET 4 83 LYS 5 84 VAL 6 85 GLY 7 86 SER 8 87 GLN 9 88 VAL 10 89 ILE 11 90 ILE 12 91 ASN 13 92 THR 14 93 SER 15 94 HIS 16 95 MET 17 96 LYS 18 97 GLY 19 98 MET 20 99 LYS 21 100 GLY 22 101 ALA 23 102 GLU 24 103 ALA 25 104 THR 26 105 VAL 27 106 THR 28 107 GLY 29 108 ALA 30 109 TYR 31 110 ASP 32 111 THR 33 112 THR 34 113 ALA 35 114 TYR 36 115 VAL 37 116 VAL 38 117 SER 39 118 TYR 40 119 THR 41 120 PRO 42 121 THR 43 122 ASN 44 123 GLY 45 124 GLY 46 125 GLN 47 126 ARG 48 127 VAL 49 128 ASP 50 129 HIS 51 130 HIS 52 131 LYS 53 132 TRP 54 133 VAL 55 134 ILE 56 135 GLN 57 136 GLU 58 137 GLU 59 138 ILE 60 139 LYS 61 140 ASP 62 141 ALA 63 142 GLY 64 143 ASP 65 144 LYS 66 145 THR 67 146 LEU 68 147 GLN 69 148 PRO 70 149 GLY 71 150 ASP 72 151 GLN 73 152 VAL 74 153 ILE 75 154 LEU 76 155 GLU 77 156 ALA 78 157 SER 79 158 HIS 80 159 MET 81 160 LYS 82 161 GLY 83 162 MET 84 163 LYS 85 164 GLY 86 165 ALA 87 166 THR 88 167 ALA 89 168 GLU 90 169 ILE 91 170 ASP 92 171 SER 93 172 ALA 94 173 GLU 95 174 LYS 96 175 THR 97 176 THR 98 177 VAL 99 178 TYR 100 179 MET 101 180 VAL 102 181 ASP 103 182 TYR 104 183 THR 105 184 SER 106 185 THR 107 186 THR 108 187 SER 109 188 GLY 110 189 GLU 111 190 LYS 112 191 VAL 113 192 LYS 114 193 ASN 115 194 HIS 116 195 LYS 117 196 TRP 118 197 VAL 119 198 THR 120 199 GLU 121 200 ASP 122 201 GLU 123 202 LEU 124 203 SER 125 204 ALA 126 205 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . 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 16942 bsCopL . . . . . PDB 2KY9 'ydhK C-terminal Domain from B.subtilis' . . . . . NESG SR518 bsCopL . . . . . stop_ save_ ############# # Ligands # ############# save_CU _Saveframe_category ligand _Mol_type "non-polymer (NON-POLYMER)" _Name_common 'COPPER (II) ION' _BMRB_code CU _PDB_code CU _Molecular_mass 63.546 _Mol_charge 2 _Mol_paramagnetic . _Mol_aromatic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons CU CU CU . 2 . ? stop_ _Mol_thiol_state . _Sequence_homology_query_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $bsCopL 'Bacillus subtilis' 1423 Bacteria . Bacillus subtilis 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 _Details $bsCopL 'recombinant technology' . Escherichia coli 'BL21 (DE3)' PET15 ; The gene fragment (residues 83-205) was cloned into a pET15 (Novagen) derivative vector yielding the construct SR518-83-205-TEV. The N-terminal Hexa-His tag was removed by TEV protease cleavage leaving additional residues (SHM) at the N-terminal of the protein ; stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_Apo-bsCopL _Saveframe_category sample _Sample_type solution _Details Apo-bsCopL loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $bsCopL 0.4 mM '[U-100% 15N]' 'TRIS pH 7.5' 10 mM 'natural abundance' 'sodium chloride' 100 mM 'natural abundance' D2O 5 % 'natural abundance' H2O 95 % 'natural abundance' stop_ save_ save_1X_Cu2+_holo-bsCopL _Saveframe_category sample _Sample_type solution _Details '1X Cu2+ holo-bsCopL' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $bsCopL 0.4 mM '[U-100% 15N]' 'Copper Sulfate' 0.4 mM 'natural abundance' 'TRIS pH 7.5' 10 mM 'natural abundance' 'sodium chloride' 100 mM 'natural abundance' D2O 5 % 'natural abundance' H2O 95 mM 'natural abundance' stop_ save_ save_2X_Cu2+_holo-bsCopL _Saveframe_category sample _Sample_type solution _Details '2X Cu2+ holo-bsCopL' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $bsCopL 0.4 mM '[U-100% 15N]' 'Copper Sulfate' 0.8 mM 'natural abundance' 'TRIS pH 7.5' 10 mM 'natural abundance' 'sodium chloride' 100 mM 'natural abundance' D2O 5 % 'natural abundance' H2O 95 % 'natural abundance' stop_ save_ save_3X_Cu2+_holo-bsCopL _Saveframe_category sample _Sample_type solution _Details '3X Cu2+ holo-bsCopL' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $bsCopL 0.4 mM '[U-100% 15N]' 'Copper Sulfate' 1.2 mM 'natural abundance' 'TRIS pH 7.5' 10 mM 'natural abundance' 'sodium chloride' 100 mM 'natural abundance' D2O 5 mM 'natural abundance' H2O 95 mM 'natural abundance' stop_ save_ save_4X_Cu2+_holo-bsCopL _Saveframe_category sample _Sample_type solution _Details '4X Cu2+ holo-bsCopL' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $bsCopL 0.4 mM '[U-100% 15N]' 'Copper Sulfate' 1.6 mM 'natural abundance' 'TRIS pH 7.5' 10 mM 'natural abundance' 'sodium chloride' 100 mM 'natural abundance' D2O 5 % 'natural abundance' H2O 95 % 'natural abundance' stop_ save_ save_6X_Cu2+_holo-bsCopL _Saveframe_category sample _Sample_type solution _Details '6X Cu2+ holo-bsCopL' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $bsCopL 0.4 mM '[U-100% 15N]' 'Copper Sulfate' 2.4 mM 'natural abundance' 'TRIS pH 7.5' 10 mM 'natural abundance' 'sodium chloride' 100 mM 'natural abundance' D2O 5 % 'natural abundance' H2O 95 % 'natural abundance' 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_NMRDraw _Saveframe_category software _Name NMRDraw _Version . loop_ _Vendor _Address _Electronic_address 'Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax' . . stop_ loop_ _Task processing stop_ _Details . save_ save_SPARKY _Saveframe_category software _Name SPARKY _Version . loop_ _Vendor _Address _Electronic_address Goddard . . stop_ loop_ _Task 'data analysis' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $Apo-bsCopL save_ save_2D_1H-15N_HSQC_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $1X_Cu2+_holo-bsCopL save_ save_2D_1H-15N_HSQC_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $2X_Cu2+_holo-bsCopL save_ save_2D_1H-15N_HSQC_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $3X_Cu2+_holo-bsCopL save_ save_2D_1H-15N_HSQC_5 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $4X_Cu2+_holo-bsCopL save_ save_2D_1H-15N_HSQC_6 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $6X_Cu2+_holo-bsCopL save_ save_2D_1H-15N_HSQC _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _BMRB_pulse_sequence_accession_number . _Details 'All NMR experiments listed below were collected using a 600 MHz spectrometer.' save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details 'Buffer used: 100 mM NaCl, 10 mM Tris pH 7.5, D2O 5%, H2O 95%' loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 100 . mM pH 7.5 . pH pressure 1 . atm temperature 273 . 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 external indirect . . . 0.25145 DSS H 1 'methyl protons' ppm 0.00 external direct . . . 1 DSS N 15 'methyl protons' ppm 0.00 external indirect . . . 0.1013 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-15N HSQC' stop_ loop_ _Sample_label $Apo-bsCopL stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name bsCopL _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 84 5 VAL H H 8.641 . . 2 84 5 VAL N N 123.376 . . 3 85 6 GLY H H 9.264 . . 4 85 6 GLY N N 116.933 . . 5 86 7 SER H H 8.161 . . 6 86 7 SER N N 117.371 . . 7 87 8 GLN H H 8.647 . . 8 87 8 GLN HE21 H 7.612 . . 9 87 8 GLN HE22 H 6.823 . . 10 87 8 GLN N N 122.019 . . 11 87 8 GLN NE2 N 112.722 . . 12 88 9 VAL H H 8.952 . . 13 88 9 VAL N N 116.895 . . 14 89 10 ILE H H 8.879 . . 15 89 10 ILE N N 120.536 . . 16 90 11 ILE H H 8.549 . . 17 90 11 ILE N N 125.302 . . 18 91 12 ASN H H 9.023 . . 19 91 12 ASN HD21 H 7.272 . . 20 91 12 ASN HD22 H 6.821 . . 21 91 12 ASN N N 126.302 . . 22 91 12 ASN ND2 N 112.224 . . 23 92 13 THR H H 6.999 . . 24 92 13 THR N N 113.124 . . 25 93 14 SER H H 7.847 . . 26 93 14 SER N N 113.900 . . 27 95 16 MET H H 7.480 . . 28 95 16 MET N N 114.970 . . 29 98 19 MET H H 7.375 . . 30 98 19 MET N N 116.407 . . 31 99 20 LYS H H 7.960 . . 32 99 20 LYS N N 119.575 . . 33 100 21 GLY H H 9.247 . . 34 100 21 GLY N N 118.056 . . 35 101 22 ALA H H 8.126 . . 36 101 22 ALA N N 123.728 . . 37 102 23 GLU H H 8.582 . . 38 102 23 GLU N N 120.251 . . 39 103 24 ALA H H 9.186 . . 40 103 24 ALA N N 127.931 . . 41 104 25 THR H H 8.941 . . 42 104 25 THR N N 115.567 . . 43 105 26 VAL H H 8.643 . . 44 105 26 VAL N N 125.464 . . 45 106 27 THR H H 9.640 . . 46 106 27 THR N N 121.518 . . 47 107 28 GLY H H 7.961 . . 48 107 28 GLY N N 110.425 . . 49 108 29 ALA H H 8.022 . . 50 108 29 ALA N N 125.309 . . 51 109 30 TYR H H 8.879 . . 52 109 30 TYR N N 120.536 . . 53 110 31 ASP H H 8.660 . . 54 110 31 ASP N N 125.856 . . 55 111 32 THR H H 7.987 . . 56 111 32 THR N N 121.401 . . 57 112 33 THR H H 8.608 . . 58 112 33 THR N N 118.564 . . 59 113 34 ALA H H 8.852 . . 60 113 34 ALA N N 129.515 . . 61 114 35 TYR H H 9.695 . . 62 114 35 TYR N N 119.484 . . 63 115 36 VAL H H 8.196 . . 64 115 36 VAL N N 124.160 . . 65 116 37 VAL H H 8.547 . . 66 116 37 VAL N N 117.880 . . 67 117 38 SER H H 8.577 . . 68 117 38 SER N N 117.611 . . 69 118 39 TYR H H 8.980 . . 70 118 39 TYR N N 120.310 . . 71 119 40 THR H H 8.436 . . 72 119 40 THR N N 120.270 . . 73 121 42 THR H H 9.713 . . 74 121 42 THR N N 114.039 . . 75 122 43 ASN H H 7.866 . . 76 122 43 ASN HD21 H 6.667 . . 77 122 43 ASN HD22 H 7.629 . . 78 122 43 ASN N N 115.955 . . 79 122 43 ASN ND2 N 111.018 . . 80 123 44 GLY H H 7.788 . . 81 123 44 GLY N N 108.090 . . 82 124 45 GLY H H 7.537 . . 83 124 45 GLY N N 105.104 . . 84 125 46 GLN H H 8.037 . . 85 125 46 GLN HE21 H 6.856 . . 86 125 46 GLN HE22 H 7.612 . . 87 125 46 GLN N N 118.300 . . 88 125 46 GLN NE2 N 112.684 . . 89 126 47 ARG H H 8.238 . . 90 126 47 ARG N N 122.199 . . 91 127 48 VAL H H 8.611 . . 92 127 48 VAL N N 129.788 . . 93 128 49 ASP H H 8.238 . . 94 128 49 ASP N N 125.708 . . 95 129 50 HIS H H 9.361 . . 96 129 50 HIS N N 122.422 . . 97 130 51 HIS H H 9.540 . . 98 130 51 HIS N N 120.234 . . 99 131 52 LYS H H 6.867 . . 100 131 52 LYS N N 128.300 . . 101 132 53 TRP H H 7.111 . . 102 132 53 TRP N N 119.296 . . 103 133 54 VAL H H 9.043 . . 104 133 54 VAL N N 115.648 . . 105 134 55 ILE H H 8.681 . . 106 134 55 ILE N N 116.293 . . 107 135 56 GLN H H 7.941 . . 108 135 56 GLN HE21 H 6.979 . . 109 135 56 GLN HE22 H 7.523 . . 110 135 56 GLN N N 122.076 . . 111 135 56 GLN NE2 N 111.276 . . 112 136 57 GLU H H 9.160 . . 113 136 57 GLU N N 113.662 . . 114 137 58 GLU H H 8.356 . . 115 137 58 GLU N N 117.902 . . 116 138 59 ILE H H 7.505 . . 117 138 59 ILE N N 122.006 . . 118 139 60 LYS H H 8.611 . . 119 139 60 LYS N N 129.788 . . 120 140 61 ASP H H 9.187 . . 121 140 61 ASP N N 121.961 . . 122 141 62 ALA H H 7.446 . . 123 141 62 ALA N N 121.090 . . 124 142 63 GLY H H 8.423 . . 125 142 63 GLY N N 104.663 . . 126 143 64 ASP H H 8.583 . . 127 143 64 ASP N N 124.873 . . 128 144 65 LYS H H 7.832 . . 129 144 65 LYS N N 120.632 . . 130 145 66 THR H H 8.562 . . 131 145 66 THR N N 120.121 . . 132 146 67 LEU H H 8.331 . . 133 146 67 LEU N N 130.800 . . 134 147 68 GLN H H 8.764 . . 135 147 68 GLN HE21 H 7.446 . . 136 147 68 GLN HE22 H 6.861 . . 137 147 68 GLN N N 118.937 . . 138 147 68 GLN NE2 N 113.257 . . 139 149 70 GLY H H 9.548 . . 140 149 70 GLY N N 115.212 . . 141 150 71 ASP H H 8.396 . . 142 150 71 ASP N N 123.355 . . 143 151 72 GLN H H 8.359 . . 144 151 72 GLN HE21 H 7.536 . . 145 151 72 GLN HE22 H 6.869 . . 146 151 72 GLN N N 119.986 . . 147 151 72 GLN NE2 N 112.614 . . 148 152 73 VAL H H 9.138 . . 149 152 73 VAL N N 117.730 . . 150 153 74 ILE H H 8.810 . . 151 153 74 ILE N N 121.168 . . 152 154 75 LEU H H 8.438 . . 153 154 75 LEU N N 123.494 . . 154 155 76 GLU H H 8.986 . . 155 155 76 GLU N N 121.459 . . 156 156 77 ALA H H 6.353 . . 157 156 77 ALA N N 121.034 . . 158 157 78 SER H H 8.081 . . 159 157 78 SER N N 109.973 . . 160 158 79 HIS H H 7.138 . . 161 158 79 HIS N N 119.930 . . 162 159 80 MET H H 7.008 . . 163 159 80 MET N N 111.004 . . 164 162 83 MET H H 7.437 . . 165 162 83 MET N N 119.078 . . 166 163 84 LYS H H 8.141 . . 167 163 84 LYS N N 119.449 . . 168 164 85 GLY H H 9.017 . . 169 164 85 GLY N N 116.599 . . 170 165 86 ALA H H 8.234 . . 171 165 86 ALA N N 124.331 . . 172 166 87 THR H H 8.516 . . 173 166 87 THR N N 116.554 . . 174 167 88 ALA H H 9.617 . . 175 167 88 ALA N N 130.934 . . 176 168 89 GLU H H 9.050 . . 177 168 89 GLU N N 119.705 . . 178 169 90 ILE H H 8.419 . . 179 169 90 ILE N N 122.663 . . 180 170 91 ASP H H 9.734 . . 181 170 91 ASP N N 129.385 . . 182 171 92 SER H H 7.834 . . 183 171 92 SER N N 110.827 . . 184 172 93 ALA H H 8.561 . . 185 172 93 ALA N N 123.866 . . 186 173 94 GLU H H 8.939 . . 187 173 94 GLU N N 121.042 . . 188 174 95 LYS H H 8.692 . . 189 174 95 LYS N N 127.630 . . 190 175 96 THR H H 8.118 . . 191 175 96 THR N N 122.328 . . 192 176 97 THR H H 8.079 . . 193 176 97 THR N N 119.077 . . 194 177 98 VAL H H 8.665 . . 195 177 98 VAL N N 120.945 . . 196 178 99 TYR H H 9.437 . . 197 178 99 TYR N N 119.154 . . 198 179 100 MET H H 8.615 . . 199 179 100 MET N N 123.625 . . 200 180 101 VAL H H 8.239 . . 201 180 101 VAL N N 114.149 . . 202 181 102 ASP H H 8.536 . . 203 181 102 ASP N N 121.963 . . 204 182 103 TYR H H 8.922 . . 205 182 103 TYR N N 118.706 . . 206 183 104 THR H H 7.963 . . 207 183 104 THR N N 117.421 . . 208 184 105 SER H H 8.751 . . 209 184 105 SER N N 121.996 . . 210 185 106 THR H H 9.926 . . 211 185 106 THR N N 121.296 . . 212 186 107 THR H H 8.072 . . 213 186 107 THR N N 112.167 . . 214 188 109 GLY H H 7.629 . . 215 188 109 GLY N N 110.970 . . 216 189 110 GLU H H 7.970 . . 217 189 110 GLU N N 120.476 . . 218 190 111 LYS H H 8.516 . . 219 190 111 LYS N N 123.688 . . 220 191 112 VAL H H 8.998 . . 221 191 112 VAL N N 130.745 . . 222 192 113 LYS H H 8.353 . . 223 192 113 LYS N N 125.547 . . 224 193 114 ASN H H 9.573 . . 225 193 114 ASN HD21 H 7.575 . . 226 193 114 ASN HD22 H 6.900 . . 227 193 114 ASN N N 119.642 . . 228 193 114 ASN ND2 N 114.066 . . 229 194 115 HIS H H 9.245 . . 230 194 115 HIS N N 118.242 . . 231 195 116 LYS H H 6.679 . . 232 195 116 LYS N N 128.370 . . 233 196 117 TRP H H 7.169 . . 234 196 117 TRP N N 118.537 . . 235 197 118 VAL H H 8.909 . . 236 197 118 VAL N N 114.344 . . 237 198 119 THR H H 9.160 . . 238 198 119 THR N N 113.662 . . 239 199 120 GLU H H 8.108 . . 240 199 120 GLU N N 118.556 . . 241 200 121 ASP H H 8.085 . . 242 200 121 ASP N N 113.212 . . 243 201 122 GLU H H 7.903 . . 244 201 122 GLU N N 118.573 . . 245 202 123 LEU H H 7.417 . . 246 202 123 LEU N N 117.259 . . 247 203 124 SER H H 8.727 . . 248 203 124 SER N N 114.296 . . 249 204 125 ALA H H 8.693 . . 250 204 125 ALA N N 126.400 . . 251 205 126 LYS H H 8.118 . . 252 205 126 LYS N N 122.328 . . stop_ save_ save_assigned_chem_shift_list_2 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-15N HSQC' stop_ loop_ _Sample_label $Apo-bsCopL stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name bsCopL _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 87 8 GLN NE2 N 112.693 . . 2 91 12 ASN ND2 N 112.234 . . 3 122 43 ASN ND2 N 110.970 . . 4 125 46 GLN NE2 N 112.722 . . 5 135 56 GLN NE2 N 111.320 . . 6 147 68 GLN NE2 N 113.254 . . 7 151 72 GLN NE2 N 112.610 . . 8 193 114 ASN ND2 N 114.063 . . stop_ save_