data_27948 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Backbone 1H, 13C, and 15N Chemical Shift Assignments for ribosomal protein bL12 S89C and with LBT tag ; _BMRB_accession_number 27948 _BMRB_flat_file_name bmr27948.str _Entry_type original _Submission_date 2019-06-17 _Accession_date 2019-06-17 _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 Wang Xiaolin . . 2 Kirkpatrick John P. . 3 Launay Helene M.M . 4 'de Simone' Alfonso . . 5 Haussinger Daniel . . 6 Dobson Christopher M. . 7 Vendruscolo Michele . . 8 Cabrita Lisa D. . 9 Waudby Christopher A. . 10 Christodoulou John . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 47 "15N chemical shifts" 47 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2019-09-25 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 27944 'bL12 dimer' stop_ _Original_release_date 2019-06-17 save_ ############################# # Citation for this entry # ############################# save_citations_1 _Saveframe_category entry_citation _Citation_full . _Citation_title ; Probing the dynamic stalk region of the ribosome using solution NMR ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 31537834 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Wang Xiaolin . . 2 Kirkpatrick John P. . 3 Launay Helene M.M . 4 'de Simone' Alfonso . . 5 Haussinger Daniel . . 6 Dobson Christopher M. . 7 Vendruscolo Michele . . 8 Cabrita Lisa D. . 9 Waudby Christopher A. . 10 Christodoulou John . . stop_ _Journal_abbreviation 'Sci. Rep.' _Journal_volume 9 _Journal_issue 1 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 13528 _Page_last 13528 _Year 2019 _Details . loop_ _Keyword NMR bL12 'lanthanide binding tag' ribosome stop_ save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'bL12 dimer' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'bL12, 1' $bL12_S89C 'bL12, 2' $bL12_S89C stop_ _System_molecular_weight 24000 _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . loop_ _Magnetic_equivalence_ID _Magnetically_equivalent_system_component 1 'bL12, 1' 1 'bL12, 2' stop_ _Database_query_date . _Details 'Complex with Lanthanide binding tag: Tm-DOTA-M8-4R4S.' save_ ######################## # Monomeric polymers # ######################## save_bL12_S89C _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common bL12_S89C _Molecular_mass . _Mol_thiol_state 'bound to LBT' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 121 _Mol_residue_sequence ; MSITKDQIIEAVAAMSVMDV VELISAMEEKFGVSAAAAVA VAAGPVEAAEEKTEFDVILK AAGANKVAVIKAVRGATGLG LKEAKDLVECAPAALKEGVS KDDAEALKKALEEAGAEVEV K ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 0 MET 2 1 SER 3 2 ILE 4 3 THR 5 4 LYS 6 5 ASP 7 6 GLN 8 7 ILE 9 8 ILE 10 9 GLU 11 10 ALA 12 11 VAL 13 12 ALA 14 13 ALA 15 14 MET 16 15 SER 17 16 VAL 18 17 MET 19 18 ASP 20 19 VAL 21 20 VAL 22 21 GLU 23 22 LEU 24 23 ILE 25 24 SER 26 25 ALA 27 26 MET 28 27 GLU 29 28 GLU 30 29 LYS 31 30 PHE 32 31 GLY 33 32 VAL 34 33 SER 35 34 ALA 36 35 ALA 37 36 ALA 38 37 ALA 39 38 VAL 40 39 ALA 41 40 VAL 42 41 ALA 43 42 ALA 44 43 GLY 45 44 PRO 46 45 VAL 47 46 GLU 48 47 ALA 49 48 ALA 50 49 GLU 51 50 GLU 52 51 LYS 53 52 THR 54 53 GLU 55 54 PHE 56 55 ASP 57 56 VAL 58 57 ILE 59 58 LEU 60 59 LYS 61 60 ALA 62 61 ALA 63 62 GLY 64 63 ALA 65 64 ASN 66 65 LYS 67 66 VAL 68 67 ALA 69 68 VAL 70 69 ILE 71 70 LYS 72 71 ALA 73 72 VAL 74 73 ARG 75 74 GLY 76 75 ALA 77 76 THR 78 77 GLY 79 78 LEU 80 79 GLY 81 80 LEU 82 81 LYS 83 82 GLU 84 83 ALA 85 84 LYS 86 85 ASP 87 86 LEU 88 87 VAL 89 88 GLU 90 89 CYS 91 90 ALA 92 91 PRO 93 92 ALA 94 93 ALA 95 94 LEU 96 95 LYS 97 96 GLU 98 97 GLY 99 98 VAL 100 99 SER 101 100 LYS 102 101 ASP 103 102 ASP 104 103 ALA 105 104 GLU 106 105 ALA 107 106 LEU 108 107 LYS 109 108 LYS 110 109 ALA 111 110 LEU 112 111 GLU 113 112 GLU 114 113 ALA 115 114 GLY 116 115 ALA 117 116 GLU 118 117 VAL 119 118 GLU 120 119 VAL 121 120 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Strain _Gene_mnemonic $bL12_S89C 'E. coli' 562 Bacteria . Escherichia coli BL21(DE3) rplL 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 $bL12_S89C 'purified from the natural source' . Escherichia coli . pET21b(+) 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 $bL12_S89C 0.9 mM '[U-13C; U-15N]' HEPES 10 mM 'natural abundance' 'magnisium chloride' 12 mM 'natural abundance' 'ammonium chloride' 30 mM 'natural abundance' beta-mercaptoethanol 2 mM 'natural abundance' DSS 0.001 % 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_CcpNMR _Saveframe_category software _Name CcpNMR _Version 'Version2.4, release 2' loop_ _Vendor _Address _Electronic_address CCPN ; Department of Biochemistry University of Cambridge 80 Tennis Court Road Old Addenbrooke's Site Cambridge CB2 1GA United Kingdom ; https://www.ccpn.ac.uk/v2-software/software/analysis stop_ loop_ _Task 'chemical shift assignment' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 700 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_1 save_ save_3D_HNCA_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCA' _Sample_label $sample_1 save_ save_3D_HN(CO)CA_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HN(CO)CA' _Sample_label $sample_1 save_ save_3D_HNCACB_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCACB' _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 42 . mM pH 7.6 . pH pressure 1 . atm temperature 298 . K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_DSS _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 internal indirect . . . 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_bL12_S89C_LBT_shift_list _Saveframe_category assigned_chemical_shifts _Details . loop_ _Software_label $CcpNMR stop_ loop_ _Experiment_label '2D 1H-15N HSQC' '3D HNCA' '3D HN(CO)CA' '3D HNCACB' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $DSS _Mol_system_component_name 'bL12, 1' _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 52 53 THR H H 8.1114 . . 2 52 53 THR N N 112.1802 . . 3 53 54 GLU H H 6.7600 . . 4 53 54 GLU N N 121.4930 . . 5 54 55 PHE H H 9.2042 . . 6 54 55 PHE N N 120.0948 . . 7 55 56 ASP H H 8.4298 . . 8 55 56 ASP N N 119.1530 . . 9 56 57 VAL H H 7.5782 . . 10 56 57 VAL N N 117.8282 . . 11 57 58 ILE H H 7.4525 . . 12 57 58 ILE N N 126.9872 . . 13 61 62 ALA H H 5.5453 . . 14 61 62 ALA N N 119.7744 . . 15 62 63 GLY H H 5.6331 . . 16 62 63 GLY N N 99.5155 . . 17 63 64 ALA H H 7.2907 . . 18 63 64 ALA N N 119.7387 . . 19 64 65 ASN H H 8.3447 . . 20 64 65 ASN N N 119.9567 . . 21 65 66 LYS H H 6.4907 . . 22 65 66 LYS N N 119.8064 . . 23 66 67 VAL H H 6.9472 . . 24 66 67 VAL N N 115.9324 . . 25 67 68 ALA H H 6.4216 . . 26 67 68 ALA N N 121.3368 . . 27 68 69 VAL H H 6.6859 . . 28 68 69 VAL N N 119.7449 . . 29 69 70 ILE H H 7.4230 . . 30 69 70 ILE N N 120.3907 . . 31 70 71 LYS H H 7.2182 . . 32 70 71 LYS N N 118.4807 . . 33 71 72 ALA H H 7.5596 . . 34 71 72 ALA N N 122.6404 . . 35 72 73 VAL H H 8.6372 . . 36 72 73 VAL N N 119.5265 . . 37 73 74 ARG H H 9.3593 . . 38 73 74 ARG N N 124.0182 . . 39 74 75 GLY H H 8.9178 . . 40 74 75 GLY N N 107.4522 . . 41 75 76 ALA H H 8.3878 . . 42 75 76 ALA N N 120.2016 . . 43 76 77 THR H H 9.2249 . . 44 76 77 THR N N 104.9013 . . 45 77 78 GLY H H 10.4428 . . 46 77 78 GLY N N 114.1791 . . 47 78 79 LEU H H 11.0783 . . 48 78 79 LEU N N 124.5196 . . 49 93 94 ALA H H 13.9288 . . 50 93 94 ALA N N 111.0532 . . 51 94 95 LEU H H 6.8420 . . 52 94 95 LEU N N 120.6947 . . 53 95 96 LYS H H 6.7510 . . 54 95 96 LYS N N 114.0865 . . 55 96 97 GLU H H 9.4306 . . 56 96 97 GLU N N 122.1697 . . 57 97 98 GLY H H 8.0853 . . 58 97 98 GLY N N 113.7392 . . 59 98 99 VAL H H 9.0631 . . 60 98 99 VAL N N 114.7475 . . 61 99 100 SER H H 9.0722 . . 62 99 100 SER N N 115.9839 . . 63 100 101 LYS H H 8.9899 . . 64 100 101 LYS N N 121.6974 . . 65 101 102 ASP H H 8.2799 . . 66 101 102 ASP N N 116.0602 . . 67 102 103 ASP H H 7.8219 . . 68 102 103 ASP N N 122.1400 . . 69 103 104 ALA H H 8.6136 . . 70 103 104 ALA N N 123.5679 . . 71 104 105 GLU H H 8.0232 . . 72 104 105 GLU N N 116.1867 . . 73 105 106 ALA H H 7.6309 . . 74 105 106 ALA N N 121.6746 . . 75 106 107 LEU H H 7.6298 . . 76 106 107 LEU N N 120.7290 . . 77 107 108 LYS H H 7.9174 . . 78 107 108 LYS N N 118.9414 . . 79 112 113 GLU H H 7.6754 . . 80 112 113 GLU N N 121.3392 . . 81 113 114 ALA H H 6.3117 . . 82 113 114 ALA N N 118.0847 . . 83 114 115 GLY H H 6.9143 . . 84 114 115 GLY N N 105.1897 . . 85 115 116 ALA H H 6.3805 . . 86 115 116 ALA N N 120.5282 . . 87 116 117 GLU H H 6.2271 . . 88 116 117 GLU N N 116.3924 . . 89 117 118 VAL H H 7.8369 . . 90 117 118 VAL N N 124.1968 . . 91 118 119 GLU H H 6.9801 . . 92 118 119 GLU N N 123.7387 . . 93 119 120 VAL H H 7.4580 . . 94 119 120 VAL N N 124.9151 . . stop_ save_