data_25783 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Astexin3 ; _BMRB_accession_number 25783 _BMRB_flat_file_name bmr25783.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" 131 "13C chemical shifts" 73 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2016-07-28 update BMRB 'update entry citation' 2015-11-09 original author 'original release' stop_ _Original_release_date 2015-11-09 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 Lisko Bozhena . . 5 Link 'A. James' . . 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 Astexin3 _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label entity $LASSO_PEPTIDE_ASTEXIN3 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_ASTEXIN3 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common LASSO_PEPTIDE_ASTEXIN3 _Molecular_mass 2542.775 _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 24 _Mol_residue_sequence ; GPTPMVGLDSVSGQYWDQHA PLAD ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 1 GLY 2 2 PRO 3 3 THR 4 4 PRO 5 5 MET 6 6 VAL 7 7 GLY 8 8 LEU 9 9 ASP 10 10 SER 11 11 VAL 12 12 SER 13 13 GLY 14 14 GLN 15 15 TYR 16 16 TRP 17 17 ASP 18 18 GLN 19 19 HIS 20 20 ALA 21 21 PRO 22 22 LEU 23 23 ALA 24 24 ASP 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_ASTEXIN3 a-proteobacteria 78587 Bacteria . Asticcacaulis excentricus 'CB 48' Astex_2447 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_ASTEXIN3 '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_ASTEXIN3 6.4 mg/mL 'natural abundance' H2O 95 % 'natural abundance' D2O 5 % '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_ ####################### # 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 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name entity _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 HA2 H 4.592 0.057 2 2 1 1 GLY HA3 H 3.843 0.041 2 3 1 1 GLY H H 8.477 0.022 1 4 1 1 GLY CA C 42.641 0.557 1 5 2 2 PRO HA H 4.755 0.047 1 6 2 2 PRO HB2 H 2.167 0.027 1 7 2 2 PRO HB3 H 2.167 0.027 1 8 2 2 PRO HG2 H 1.843 0.049 2 9 2 2 PRO HG3 H 2.005 0.027 2 10 2 2 PRO HD2 H 3.453 0.030 2 11 2 2 PRO HD3 H 3.575 0.024 2 12 2 2 PRO CB C 27.521 0.605 1 13 2 2 PRO CG C 24.029 0.488 1 14 2 2 PRO CD C 46.505 0.537 1 15 3 3 THR H H 7.697 0.027 1 16 3 3 THR HA H 4.661 0.065 1 17 3 3 THR HB H 4.328 0.051 1 18 3 3 THR HG2 H 1.078 0.048 1 19 3 3 THR CA C 58.119 0.417 1 20 3 3 THR CB C 67.067 0.435 1 21 3 3 THR CG2 C 18.203 0.557 1 22 4 4 PRO HA H 4.163 0.020 1 23 4 4 PRO HB2 H 1.869 0.034 2 24 4 4 PRO HB3 H 1.934 0.033 2 25 4 4 PRO HG2 H 1.831 0.041 1 26 4 4 PRO HG3 H 1.831 0.041 1 27 4 4 PRO HD2 H 3.615 0.034 2 28 4 4 PRO HD3 H 3.664 0.028 2 29 4 4 PRO CA C 60.219 0.555 1 30 4 4 PRO CB C 28.224 0.560 1 31 4 4 PRO CG C 27.519 0.536 1 32 4 4 PRO CD C 47.911 0.547 1 33 5 5 MET H H 7.527 0.024 1 34 5 5 MET HA H 4.630 0.063 1 35 5 5 MET HB2 H 2.043 0.059 1 36 5 5 MET HB3 H 2.043 0.059 1 37 5 5 MET HG2 H 2.461 0.042 1 38 5 5 MET HG3 H 2.461 0.042 1 39 5 5 MET HE H 1.942 0.029 1 40 5 5 MET CB C 31.723 0.539 1 41 5 5 MET CG C 29.444 0.578 1 42 5 5 MET CE C 14.166 0.734 1 43 6 6 VAL H H 8.442 0.022 1 44 6 6 VAL HA H 3.842 0.034 1 45 6 6 VAL HB H 1.818 0.041 1 46 6 6 VAL HG1 H 0.858 0.052 2 47 6 6 VAL HG2 H 0.758 0.049 2 48 6 6 VAL CA C 61.277 0.516 1 49 6 6 VAL CB C 29.281 0.655 1 50 6 6 VAL CG1 C 18.945 0.477 2 51 6 6 VAL CG2 C 18.373 0.438 2 52 7 7 GLY H H 8.267 0.016 1 53 7 7 GLY HA2 H 3.408 0.041 2 54 7 7 GLY HA3 H 2.876 0.040 2 55 7 7 GLY CA C 42.283 0.395 1 56 8 8 LEU H H 6.472 0.029 1 57 8 8 LEU HA H 4.086 0.027 1 58 8 8 LEU HB2 H 1.332 0.038 2 59 8 8 LEU HB3 H 1.286 0.036 2 60 8 8 LEU HG H 1.192 0.040 1 61 8 8 LEU HD1 H 0.746 0.030 2 62 8 8 LEU HD2 H 0.664 0.040 2 63 8 8 LEU CA C 52.134 0.507 1 64 8 8 LEU CB C 39.958 0.492 1 65 8 8 LEU CG C 24.003 0.578 1 66 8 8 LEU CD1 C 21.736 0.449 1 67 8 8 LEU CD2 C 21.188 0.495 2 68 9 9 ASP H H 8.401 0.024 1 69 9 9 ASP HA H 4.670 0.047 1 70 9 9 ASP HB2 H 3.327 0.055 2 71 9 9 ASP HB3 H 2.305 0.053 2 72 9 9 ASP CB C 39.699 0.492 1 73 10 10 SER H H 9.124 0.019 1 74 10 10 SER HA H 4.254 0.028 1 75 10 10 SER HB2 H 3.925 0.042 2 76 10 10 SER HB3 H 3.808 0.047 2 77 10 10 SER CA C 57.056 0.633 1 78 10 10 SER CB C 60.570 0.541 1 79 11 11 VAL H H 8.901 0.037 1 80 11 11 VAL HA H 4.084 0.040 1 81 11 11 VAL HB H 2.052 0.069 1 82 11 11 VAL HG1 H 0.839 0.054 1 83 11 11 VAL HG2 H 0.839 0.054 1 84 11 11 VAL CA C 61.750 0.578 1 85 11 11 VAL CB C 30.568 0.447 1 86 11 11 VAL CG1 C 18.378 0.465 2 87 11 11 VAL CG2 C 18.378 0.465 2 88 12 12 SER H H 7.699 0.017 1 89 12 12 SER HA H 4.436 0.033 1 90 12 12 SER HB2 H 3.761 0.031 2 91 12 12 SER HB3 H 3.723 0.025 2 92 12 12 SER CA C 55.999 0.500 1 93 12 12 SER CB C 61.279 0.621 1 94 13 13 GLY H H 8.537 0.027 1 95 13 13 GLY HA2 H 4.160 0.051 2 96 13 13 GLY HA3 H 3.833 0.057 2 97 13 13 GLY CA C 42.303 0.481 1 98 14 14 GLN H H 8.150 0.020 1 99 14 14 GLN HA H 4.261 0.030 1 100 14 14 GLN HB2 H 1.554 0.051 1 101 14 14 GLN HB3 H 1.554 0.051 1 102 14 14 GLN HG2 H 1.862 0.035 2 103 14 14 GLN HG3 H 1.767 0.037 2 104 14 14 GLN HE21 H 6.682 0.055 2 105 14 14 GLN HE22 H 7.139 0.031 2 106 14 14 GLN CA C 53.044 0.495 1 107 14 14 GLN CB C 26.805 0.596 1 108 14 14 GLN CG C 30.684 0.581 1 109 15 15 TYR H H 8.232 0.024 1 110 15 15 TYR HA H 4.934 0.062 1 111 15 15 TYR HB2 H 3.293 0.068 2 112 15 15 TYR HB3 H 2.937 0.067 2 113 15 15 TYR HD1 H 6.841 0.032 3 114 15 15 TYR HD2 H 6.841 0.032 3 115 15 15 TYR HE1 H 6.617 0.021 1 116 15 15 TYR HE2 H 6.617 0.021 1 117 15 15 TYR CB C 37.370 0.526 1 118 15 15 TYR CD1 C 130.557 0.562 2 119 15 15 TYR CD2 C 130.557 0.562 2 120 15 15 TYR CE1 C 114.908 0.552 2 121 15 15 TYR CE2 C 114.908 0.552 2 122 16 16 TRP H H 8.641 0.028 1 123 16 16 TRP HA H 4.197 0.032 1 124 16 16 TRP HB2 H 3.215 0.048 2 125 16 16 TRP HB3 H 2.803 0.053 2 126 16 16 TRP HD1 H 7.232 0.028 1 127 16 16 TRP HE1 H 10.081 0.021 1 128 16 16 TRP HE3 H 7.604 0.024 1 129 16 16 TRP HZ2 H 7.389 0.028 1 130 16 16 TRP HZ3 H 7.096 0.079 1 131 16 16 TRP HH2 H 7.153 0.062 1 132 16 16 TRP CA C 54.464 0.518 1 133 16 16 TRP CB C 28.028 0.473 1 134 16 16 TRP CD1 C 124.782 0.509 1 135 16 16 TRP CE3 C 118.598 0.443 1 136 16 16 TRP CZ2 C 112.262 0.417 1 137 16 16 TRP CZ3 C 119.484 0.443 1 138 16 16 TRP CH2 C 122.465 0.463 1 139 17 17 ASP H H 7.381 0.026 1 140 17 17 ASP HA H 4.295 0.031 1 141 17 17 ASP HB2 H 2.705 0.061 1 142 17 17 ASP HB3 H 2.705 0.061 1 143 17 17 ASP CA C 50.728 0.559 1 144 17 17 ASP CB C 37.014 0.617 1 145 18 18 GLN H H 7.690 0.025 1 146 18 18 GLN HA H 3.421 0.027 1 147 18 18 GLN HB2 H 1.689 0.046 2 148 18 18 GLN HB3 H 1.566 0.040 2 149 18 18 GLN HG2 H 1.948 0.050 1 150 18 18 GLN HG3 H 1.948 0.050 1 151 18 18 GLN HE21 H 6.787 0.030 2 152 18 18 GLN HE22 H 7.315 0.018 2 153 18 18 GLN CA C 53.543 0.559 1 154 18 18 GLN CB C 26.467 0.547 1 155 18 18 GLN CG C 31.126 0.607 1 156 19 19 HIS H H 8.232 0.018 1 157 19 19 HIS HA H 4.475 0.034 1 158 19 19 HIS HB2 H 3.068 0.041 2 159 19 19 HIS HB3 H 2.974 0.039 2 160 19 19 HIS HD2 H 7.079 0.011 1 161 19 19 HIS HE1 H 8.347 0.026 1 162 19 19 HIS CA C 52.135 0.513 1 163 19 19 HIS CB C 26.464 0.536 1 164 19 19 HIS CD2 C 117.402 0.483 1 165 19 19 HIS CE1 C 133.370 0.549 1 166 20 20 ALA H H 8.285 0.018 1 167 20 20 ALA HA H 4.421 0.034 1 168 20 20 ALA HB H 1.229 0.051 1 169 20 20 ALA CA C 47.918 0.518 1 170 20 20 ALA CB C 15.213 0.596 1 171 21 21 PRO HA H 4.291 0.040 1 172 21 21 PRO HB2 H 1.771 0.038 2 173 21 21 PRO HB3 H 2.165 0.056 2 174 21 21 PRO HG2 H 1.901 0.036 1 175 21 21 PRO HG3 H 1.901 0.036 1 176 21 21 PRO HD2 H 3.497 0.025 2 177 21 21 PRO HD3 H 3.660 0.026 2 178 21 21 PRO CA C 60.368 0.518 1 179 21 21 PRO CB C 29.244 0.526 1 180 21 21 PRO CG C 24.683 0.557 1 181 21 21 PRO CD C 47.629 0.481 1 182 22 22 LEU H H 8.178 0.017 1 183 22 22 LEU HA H 4.144 0.036 1 184 22 22 LEU HB2 H 1.439 0.029 1 185 22 22 LEU HB3 H 1.439 0.029 1 186 22 22 LEU HG H 1.490 0.031 1 187 22 22 LEU HD1 H 0.760 0.029 2 188 22 22 LEU HD2 H 0.694 0.041 2 189 22 22 LEU CA C 52.327 0.596 1 190 22 22 LEU CB C 39.612 0.548 1 191 22 22 LEU CG C 24.286 0.545 1 192 22 22 LEU CD1 C 22.041 0.419 2 193 22 22 LEU CD2 C 20.700 0.408 2 194 23 23 ALA H H 8.121 0.013 1 195 23 23 ALA HA H 4.200 0.045 1 196 23 23 ALA HB H 1.239 0.041 1 197 23 23 ALA CA C 49.479 0.536 1 198 23 23 ALA CB C 16.576 0.605 1 199 24 24 ASP H H 8.152 0.034 1 200 24 24 ASP HA H 4.573 0.028 1 201 24 24 ASP HB2 H 2.793 0.063 1 202 24 24 ASP HB3 H 2.793 0.063 1 203 24 24 ASP CA C 49.529 0.500 1 204 24 24 ASP CB C 35.814 0.674 1 stop_ save_