data_25782 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Astexin2-dC4 ; _BMRB_accession_number 25782 _BMRB_flat_file_name bmr25782.str _Entry_type original _Submission_date 2015-08-28 _Accession_date 2015-08-28 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details 'LASSO PEPTIDE, ANTIMICROBIAL PEPTIDE' loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Link 'A. James' . . 2 Maksimov Mikhail O. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 106 "13C chemical shifts" 59 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2016-07-28 update BMRB 'update entry citation' 2015-11-18 update author 'update shifts, etc.' 2015-11-17 original author 'original release' stop_ loop_ _Related_BMRB_accession_number _Relationship 25783 Astexin3 stop_ _Original_release_date 2015-11-17 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Elucidating the Specificity Determinants of the AtxE2 Lasso Peptide Isopeptidase ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 26534965 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Maksimov Mikhail O. . 2 Koos Joseph D. . 3 Zong Chuhan . . 4 Link 'A. James' . . 5 Lisko Bozhena . . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_volume 290 _Journal_issue 52 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 30806 _Page_last 30812 _Year 2015 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'Lasso peptide Astexin2-dC4' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'Lasso peptide Astexin2-dC4' $LASSO_PEPTIDE_ASTEXIN2-dC4 stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_LASSO_PEPTIDE_ASTEXIN2-dC4 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common LASSO_PEPTIDE_ASTEXIN2-dC4 _Molecular_mass 2134.348 _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 20 _Mol_residue_sequence ; GLTQIQALDSVSGQFRDQLG ; loop_ _Residue_seq_code _Residue_label 1 GLY 2 LEU 3 THR 4 GLN 5 ILE 6 GLN 7 ALA 8 LEU 9 ASP 10 SER 11 VAL 12 SER 13 GLY 14 GLN 15 PHE 16 ARG 17 ASP 18 GLN 19 LEU 20 GLY 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 $LASSO_PEPTIDE_ASTEXIN2-dC4 a-proteobacteria 78587 Bacteria . Asticcacaulis excentricus 'CB 48' Astex_2448 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 $LASSO_PEPTIDE_ASTEXIN2-dC4 'recombinant technology' . Escherichia coli BL21 pASK75 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 $LASSO_PEPTIDE_ASTEXIN2-dC4 2.7 mg/mL 'natural abundance' stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $LASSO_PEPTIDE_ASTEXIN2-dC4 4.9 mM 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_CYANA _Saveframe_category software _Name CYANA _Version 2.1 loop_ _Vendor _Address _Electronic_address 'Guntert, Mumenthaler and Wuthrich' . . stop_ loop_ _Task 'structure solution' stop_ _Details . save_ save_Mestrenova _Saveframe_category software _Name Mestrenova _Version 9.0.0 loop_ _Vendor _Address _Electronic_address 'Mestrelab Research' . . stop_ loop_ _Task 'data analysis' stop_ _Details . save_ save_GROMACS _Saveframe_category software _Name GROMACS _Version 4.6.5 loop_ _Vendor _Address _Electronic_address 'Hess, Kutzner, van der Spoel and Lindahl' . . stop_ loop_ _Task refinement stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 800 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_DQF-COSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D DQF-COSY' _Sample_label $sample_1 save_ save_2D_1H-1H_NOESY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $sample_1 save_ save_2D_1H-1H_TOCSY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label $sample_1 save_ save_2D_1H-13C_HSQC_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HSQC' _Sample_label $sample_1 save_ save_2D_1H-13C_HSQC_5 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HSQC' _Sample_label $sample_2 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pressure 1 . atm temperature 295 . 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 water C 13 protons ppm 4.7 internal indirect . . . 0.251467875 water H 1 protons ppm 4.7 internal direct . . . 1.000000000 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_ _Software_label $Mestrenova stop_ loop_ _Experiment_label '2D DQF-COSY' '2D 1H-1H NOESY' '2D 1H-1H TOCSY' '2D 1H-13C HSQC' stop_ loop_ _Sample_label $sample_1 $sample_2 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name 'Lasso peptide Astexin2-dC4' _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 1 1 GLY H H 8.951 0.032 1 2 1 1 GLY HA2 H 4.718 0.045 2 3 1 1 GLY HA3 H 3.749 0.048 2 4 1 1 GLY CA C 43.022 0.424 1 5 2 2 LEU H H 8.218 0.025 1 6 2 2 LEU HA H 4.654 0.033 1 7 2 2 LEU HB2 H 1.584 0.033 2 8 2 2 LEU HB3 H 1.493 0.043 2 9 2 2 LEU HG H 1.584 0.029 1 10 2 2 LEU HD1 H 0.830 0.032 2 11 2 2 LEU HD2 H 0.740 0.029 2 12 2 2 LEU CB C 38.069 0.454 1 13 2 2 LEU CG C 24.360 0.439 1 14 2 2 LEU CD1 C 22.475 0.479 2 15 2 2 LEU CD2 C 20.500 0.424 2 16 3 3 THR H H 7.153 0.025 1 17 3 3 THR HA H 4.427 0.032 1 18 3 3 THR HB H 4.274 0.044 1 19 3 3 THR HG2 H 1.021 0.033 1 20 3 3 THR CA C 58.900 0.378 1 21 3 3 THR CB C 69.080 0.575 1 22 3 3 THR CG2 C 18.137 0.502 1 23 4 4 GLN H H 8.138 0.021 1 24 4 4 GLN HA H 3.801 0.029 1 25 4 4 GLN HB2 H 2.104 0.037 2 26 4 4 GLN HB3 H 1.522 0.041 2 27 4 4 GLN HG2 H 1.983 0.025 2 28 4 4 GLN HG3 H 1.921 0.032 2 29 4 4 GLN CA C 51.903 0.366 1 30 4 4 GLN CB C 26.037 0.475 1 31 4 4 GLN CG C 31.336 0.642 1 32 5 5 ILE H H 7.613 0.030 1 33 5 5 ILE HA H 4.097 0.028 1 34 5 5 ILE HB H 1.632 0.046 1 35 5 5 ILE HG12 H 1.030 0.037 2 36 5 5 ILE HG13 H 1.450 0.045 2 37 5 5 ILE HG2 H 0.770 0.036 1 38 5 5 ILE HD1 H 0.770 0.036 1 39 5 5 ILE CB C 36.889 0.523 1 40 5 5 ILE CG1 C 24.696 0.498 1 41 5 5 ILE CG2 C 14.453 0.671 1 42 5 5 ILE CD1 C 10.317 0.702 1 43 6 6 GLN H H 8.982 0.034 1 44 6 6 GLN HA H 4.273 0.036 1 45 6 6 GLN HB2 H 1.936 0.051 1 46 6 6 GLN HB3 H 1.936 0.051 1 47 6 6 GLN HG2 H 2.324 0.043 2 48 6 6 GLN HG3 H 2.121 0.045 2 49 6 6 GLN CA C 55.567 0.266 1 50 6 6 GLN CB C 25.810 0.533 1 51 6 6 GLN CG C 31.497 0.404 1 52 7 7 ALA H H 8.226 0.015 1 53 7 7 ALA HA H 4.053 0.039 1 54 7 7 ALA HB H 1.234 0.038 1 55 7 7 ALA CA C 49.626 0.469 1 56 7 7 ALA CB C 17.129 0.533 1 57 8 8 LEU H H 7.617 0.028 1 58 8 8 LEU HA H 3.987 0.027 1 59 8 8 LEU HB2 H 1.414 0.040 2 60 8 8 LEU HB3 H 1.338 0.043 2 61 8 8 LEU HG H 1.414 0.054 1 62 8 8 LEU HD1 H 0.790 0.036 2 63 8 8 LEU HD2 H 0.710 0.025 2 64 8 8 LEU CA C 53.182 0.305 1 65 8 8 LEU CB C 39.472 0.617 1 66 8 8 LEU CG C 24.004 0.512 1 67 8 8 LEU CD1 C 21.402 0.439 1 68 8 8 LEU CD2 C 21.940 0.408 1 69 9 9 ASP H H 7.375 0.033 1 70 9 9 ASP HA H 4.364 0.029 1 71 9 9 ASP HB2 H 3.239 0.044 2 72 9 9 ASP HB3 H 2.046 0.047 2 73 9 9 ASP CA C 51.417 0.282 1 74 9 9 ASP CB C 41.032 0.423 1 75 10 10 SER H H 9.635 0.039 1 76 10 10 SER HA H 4.224 0.022 1 77 10 10 SER HB2 H 3.927 0.040 2 78 10 10 SER HB3 H 3.851 0.037 2 79 10 10 SER CA C 58.245 0.472 1 80 10 10 SER CB C 59.928 0.434 1 81 11 11 VAL H H 8.955 0.033 1 82 11 11 VAL HA H 4.044 0.041 1 83 11 11 VAL HB H 2.054 0.038 1 84 11 11 VAL HG1 H 0.841 0.035 1 85 11 11 VAL HG2 H 0.841 0.035 1 86 11 11 VAL CA C 62.108 0.378 1 87 11 11 VAL CB C 30.404 0.585 1 88 11 11 VAL CG1 C 18.638 0.448 2 89 11 11 VAL CG2 C 18.347 0.563 2 90 12 12 SER H H 7.700 0.020 1 91 12 12 SER HA H 4.483 0.042 1 92 12 12 SER HB2 H 3.741 0.036 2 93 12 12 SER HB3 H 3.591 0.039 2 94 12 12 SER CA C 55.522 0.545 1 95 12 12 SER CB C 61.278 0.576 1 96 13 13 GLY H H 8.790 0.036 1 97 13 13 GLY HA2 H 4.269 0.040 2 98 13 13 GLY HA3 H 3.825 0.051 2 99 13 13 GLY CA C 42.286 0.391 1 100 14 14 GLN H H 8.160 0.022 1 101 14 14 GLN HA H 4.301 0.035 1 102 14 14 GLN HB2 H 1.353 0.038 1 103 14 14 GLN HB3 H 1.353 0.038 1 104 14 14 GLN HG2 H 1.857 0.031 2 105 14 14 GLN HG3 H 1.790 0.032 2 106 14 14 GLN CA C 52.506 0.523 1 107 14 14 GLN CB C 26.373 0.430 1 108 14 14 GLN CG C 30.696 0.518 1 109 15 15 PHE H H 8.170 0.024 1 110 15 15 PHE HA H 5.397 0.044 1 111 15 15 PHE HB2 H 2.958 0.038 2 112 15 15 PHE HB3 H 2.779 0.042 2 113 15 15 PHE HD1 H 6.937 0.025 1 114 15 15 PHE HD2 H 6.937 0.025 1 115 15 15 PHE HE1 H 7.154 0.022 1 116 15 15 PHE HE2 H 7.154 0.022 1 117 15 15 PHE HZ H 7.220 0.043 1 118 15 15 PHE CA C 53.504 0.326 1 119 15 15 PHE CB C 39.035 0.502 1 120 15 15 PHE CD1 C 129.610 0.762 3 121 15 15 PHE CD2 C 129.610 0.762 3 122 15 15 PHE CE1 C 128.088 0.511 3 123 15 15 PHE CE2 C 128.088 0.511 3 124 15 15 PHE CZ C 128.797 0.527 1 125 16 16 ARG H H 8.904 0.029 1 126 16 16 ARG HA H 3.592 0.030 1 127 16 16 ARG HB2 H 1.644 0.032 2 128 16 16 ARG HB3 H 1.367 0.035 2 129 16 16 ARG HG2 H 1.994 0.042 2 130 16 16 ARG HG3 H 1.602 0.033 2 131 16 16 ARG HD2 H 3.058 0.030 2 132 16 16 ARG HD3 H 2.900 0.018 2 133 16 16 ARG HE H 6.953 0.022 1 134 16 16 ARG CA C 53.161 0.497 1 135 16 16 ARG CB C 29.042 0.413 1 136 16 16 ARG CG C 24.266 0.397 1 137 16 16 ARG CD C 41.353 0.523 1 138 17 17 ASP H H 8.505 0.022 1 139 17 17 ASP HA H 4.331 0.034 1 140 17 17 ASP HB2 H 2.814 0.036 2 141 17 17 ASP HB3 H 2.542 0.048 2 142 17 17 ASP CB C 36.517 0.512 1 143 18 18 GLN H H 8.606 0.020 1 144 18 18 GLN HA H 3.889 0.030 1 145 18 18 GLN HB2 H 1.944 0.055 2 146 18 18 GLN HB3 H 1.830 0.044 2 147 18 18 GLN HG2 H 2.223 0.035 1 148 18 18 GLN HG3 H 2.223 0.035 1 149 18 18 GLN HE21 H 6.850 0.040 2 150 18 18 GLN HE22 H 7.480 0.022 2 151 18 18 GLN CA C 54.246 0.491 1 152 18 18 GLN CB C 26.503 0.664 1 153 18 18 GLN CG C 31.123 0.554 1 154 19 19 LEU H H 8.305 0.017 1 155 19 19 LEU HA H 4.273 0.034 1 156 19 19 LEU HB2 H 1.523 0.035 2 157 19 19 LEU HB3 H 1.493 0.031 2 158 19 19 LEU HG H 1.493 0.025 1 159 19 19 LEU HD1 H 0.800 0.029 2 160 19 19 LEU HD2 H 0.760 0.021 2 161 19 19 LEU CA C 52.367 0.403 1 162 19 19 LEU CB C 39.640 0.513 1 163 19 19 LEU CG C 24.274 0.471 1 164 19 19 LEU CD1 C 22.106 0.493 2 165 19 19 LEU CD2 C 20.827 0.292 2 stop_ save_