data_27501 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H and 15N Chemical Shift Assignments for SP6 variant of SPINK1 ; _BMRB_accession_number 27501 _BMRB_flat_file_name bmr27501.str _Entry_type original _Submission_date 2018-06-02 _Accession_date 2018-06-02 _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 Sebak Fanni . . 2 Bodor Andrea . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 177 "15N chemical shifts" 45 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2019-02-07 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 27500 'SP5 variant of SPINK1' stop_ _Original_release_date 2018-06-04 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Directed Evolution of Canonical Loops and Their Swapping between Unrelated Serine Proteinase Inhibitors Disprove the Interscaffolding Additivity Model ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 30543823 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Boros Eszter . . 2 Sebak Fanni . . 3 Heja David . . 4 Szakacs David . . 5 Zboray Katalin . . 6 Schlosser Gitta . . 7 Micsonai Andras . . 8 Kardos Jozsef . . 9 Bodor Andrea . . 10 Pal Gabor . . stop_ _Journal_abbreviation 'J. Mol. Biol.' _Journal_name_full 'Journal of molecular biology' _Journal_volume 431 _Journal_issue 3 _Journal_ISSN 1089-8638 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 557 _Page_last 575 _Year 2019 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name SPINK1 _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label SPINK1 $SPINK1 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_SPINK1 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common SPINK1 _Molecular_mass . _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 59 _Mol_residue_sequence ; GSGDSLGREAKCYNELNDCT MKMAKVCGTDGNTYPNECVL CFENRKRQTSILIQKSGPC ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 -2 GLY 2 -1 SER 3 0 GLY 4 1 ASP 5 2 SER 6 3 LEU 7 4 GLY 8 5 ARG 9 6 GLU 10 7 ALA 11 8 LYS 12 9 CYS 13 10 TYR 14 11 ASN 15 12 GLU 16 13 LEU 17 14 ASN 18 15 ASP 19 16 CYS 20 17 THR 21 18 MET 22 19 LYS 23 20 MET 24 21 ALA 25 22 LYS 26 23 VAL 27 24 CYS 28 25 GLY 29 26 THR 30 27 ASP 31 28 GLY 32 29 ASN 33 30 THR 34 31 TYR 35 32 PRO 36 33 ASN 37 34 GLU 38 35 CYS 39 36 VAL 40 37 LEU 41 38 CYS 42 39 PHE 43 40 GLU 44 41 ASN 45 42 ARG 46 43 LYS 47 44 ARG 48 45 GLN 49 46 THR 50 47 SER 51 48 ILE 52 49 LEU 53 50 ILE 54 51 GLN 55 52 LYS 56 53 SER 57 54 GLY 58 55 PRO 59 56 CYS 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 $SPINK1 Human 9606 Eukaryota Metazoa Homo sapiens 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 $SPINK1 'recombinant technology' . Escherichia coli . M13KO7 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 $SPINK1 1 mM '[U-100% 15N]' MES 10 mM 'natural abundance' 'sodium chloride' 50 mM 'natural abundance' DSS 5 uL 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_TOPSPIN _Saveframe_category software _Name TOPSPIN _Version . loop_ _Vendor _Address _Electronic_address 'Bruker Biospin' . . stop_ loop_ _Task processing stop_ _Details . save_ save_CARA _Saveframe_category software _Name CARA _Version . loop_ _Vendor _Address _Electronic_address 'Keller and Wuthrich' . . 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_1H-15N_TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-15N TOCSY' _Sample_label $sample_1 save_ save_3D_1H-15N_NOESY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-15N NOESY' _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 pH 3 . pH pressure 1 . atm temperature 298 . 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 H 1 'methyl protons' ppm 0.00 internal direct . . . 1.000000000 DSS N 15 'methyl protons' ppm 0.00 na indirect . . . 0.101329118 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' '3D 1H-15N TOCSY' '3D 1H-15N NOESY' 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 SPINK1 _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 4 ASP H H 8.132 0.02 1 2 1 4 ASP HA H 4.617 0.02 1 3 1 4 ASP HB2 H 2.861 0.02 2 4 1 4 ASP HB3 H 2.861 0.02 2 5 1 4 ASP N N 118.945 0.3 1 6 2 5 SER H H 8.339 0.02 1 7 2 5 SER HA H 4.420 0.02 1 8 2 5 SER HB2 H 3.909 0.02 2 9 2 5 SER HB3 H 3.909 0.02 2 10 2 5 SER N N 116.367 0.3 1 11 3 6 LEU H H 8.270 0.02 1 12 3 6 LEU HA H 4.381 0.02 1 13 3 6 LEU HB2 H 1.668 0.02 2 14 3 6 LEU HB3 H 1.668 0.02 2 15 3 6 LEU HG H 1.578 0.02 1 16 3 6 LEU HD1 H 0.845 0.02 2 17 3 6 LEU HD2 H 0.845 0.02 2 18 3 6 LEU N N 123.805 0.3 1 19 4 7 GLY H H 8.100 0.02 1 20 4 7 GLY HA2 H 3.924 0.02 2 21 4 7 GLY HA3 H 3.924 0.02 2 22 4 7 GLY N N 107.913 0.3 1 23 5 8 ARG H H 8.267 0.02 1 24 5 8 ARG HA H 4.682 0.02 1 25 5 8 ARG HB2 H 1.773 0.02 2 26 5 8 ARG HB3 H 1.773 0.02 2 27 5 8 ARG N N 119.013 0.3 1 28 6 9 GLU H H 8.529 0.02 1 29 6 9 GLU HA H 4.276 0.02 1 30 6 9 GLU HB2 H 1.891 0.02 2 31 6 9 GLU HB3 H 1.891 0.02 2 32 6 9 GLU HG2 H 2.271 0.02 2 33 6 9 GLU HG3 H 2.271 0.02 2 34 6 9 GLU N N 123.308 0.3 1 35 7 10 ALA H H 8.727 0.02 1 36 7 10 ALA HA H 4.313 0.02 1 37 7 10 ALA HB H 1.310 0.02 1 38 7 10 ALA N N 126.960 0.3 1 39 11 14 ASN H H 8.408 0.02 1 40 11 14 ASN HA H 4.620 0.02 1 41 11 14 ASN HB2 H 2.835 0.02 2 42 11 14 ASN HB3 H 2.835 0.02 2 43 11 14 ASN N N 118.355 0.3 1 44 12 15 GLU H H 8.022 0.02 1 45 12 15 GLU HA H 4.430 0.02 1 46 12 15 GLU HB2 H 2.417 0.02 1 47 12 15 GLU HB3 H 1.987 0.02 1 48 12 15 GLU N N 118.323 0.3 1 49 13 16 LEU H H 8.189 0.02 1 50 13 16 LEU HA H 4.153 0.02 1 51 13 16 LEU HB2 H 1.567 0.02 2 52 13 16 LEU HB3 H 1.567 0.02 2 53 13 16 LEU N N 122.210 0.3 1 54 14 17 ASN H H 8.337 0.02 1 55 14 17 ASN HA H 4.537 0.02 1 56 14 17 ASN HB2 H 2.748 0.02 2 57 14 17 ASN HB3 H 2.748 0.02 2 58 14 17 ASN N N 116.855 0.3 1 59 15 18 ASP H H 8.236 0.02 1 60 15 18 ASP HA H 4.617 0.02 1 61 15 18 ASP HB2 H 2.821 0.02 2 62 15 18 ASP HB3 H 2.821 0.02 2 63 15 18 ASP N N 116.984 0.3 1 64 16 19 CYS H H 8.238 0.02 1 65 16 19 CYS HA H 4.692 0.02 1 66 16 19 CYS HB2 H 2.783 0.02 2 67 16 19 CYS HB3 H 2.783 0.02 2 68 16 19 CYS N N 118.335 0.3 1 69 17 20 THR H H 8.182 0.02 1 70 17 20 THR HA H 4.331 0.02 1 71 17 20 THR N N 115.377 0.3 1 72 21 24 ALA H H 8.245 0.02 1 73 21 24 ALA HA H 4.251 0.02 1 74 21 24 ALA HB H 1.379 0.02 1 75 21 24 ALA N N 128.182 0.3 1 76 23 26 VAL H H 8.071 0.02 1 77 23 26 VAL HA H 4.818 0.02 1 78 23 26 VAL HB H 1.505 0.02 1 79 23 26 VAL HG1 H 0.619 0.02 2 80 23 26 VAL HG2 H 0.619 0.02 2 81 23 26 VAL N N 109.868 0.3 1 82 24 27 CYS H H 8.516 0.02 1 83 24 27 CYS HA H 5.377 0.02 1 84 24 27 CYS HB2 H 2.913 0.02 2 85 24 27 CYS HB3 H 2.808 0.02 2 86 24 27 CYS N N 121.864 0.3 1 87 25 28 GLY H H 9.765 0.02 1 88 25 28 GLY HA2 H 5.164 0.02 1 89 25 28 GLY HA3 H 4.253 0.02 1 90 25 28 GLY N N 116.023 0.3 1 91 26 29 THR H H 9.056 0.02 1 92 26 29 THR HA H 4.197 0.02 1 93 26 29 THR HB H 3.455 0.02 1 94 26 29 THR HG2 H 1.170 0.02 1 95 26 29 THR N N 114.034 0.3 1 96 27 30 ASP H H 8.456 0.02 1 97 27 30 ASP HA H 4.538 0.02 1 98 27 30 ASP HB2 H 3.044 0.02 1 99 27 30 ASP HB3 H 2.743 0.02 1 100 27 30 ASP N N 118.539 0.3 1 101 28 31 GLY H H 8.328 0.02 1 102 28 31 GLY HA2 H 4.105 0.02 1 103 28 31 GLY HA3 H 3.752 0.02 1 104 28 31 GLY N N 108.595 0.3 1 105 29 32 ASN H H 7.904 0.02 1 106 29 32 ASN HA H 4.957 0.02 1 107 29 32 ASN HB2 H 2.847 0.02 1 108 29 32 ASN HB3 H 2.166 0.02 1 109 29 32 ASN N N 118.307 0.3 1 110 30 33 THR H H 8.580 0.02 1 111 30 33 THR HA H 4.984 0.02 1 112 30 33 THR HB H 3.988 0.02 1 113 30 33 THR HG2 H 1.164 0.02 1 114 30 33 THR N N 120.879 0.3 1 115 31 34 TYR H H 9.676 0.02 1 116 31 34 TYR HA H 4.800 0.02 1 117 31 34 TYR HB2 H 2.671 0.02 2 118 31 34 TYR HB3 H 2.671 0.02 2 119 31 34 TYR N N 127.644 0.3 1 120 33 36 ASN H H 6.989 0.02 1 121 33 36 ASN HA H 5.023 0.02 1 122 33 36 ASN N N 129.097 0.3 1 123 34 37 GLU H H 9.631 0.02 1 124 34 37 GLU HA H 3.896 0.02 1 125 34 37 GLU HB2 H 2.093 0.02 2 126 34 37 GLU HB3 H 2.093 0.02 2 127 34 37 GLU N N 117.197 0.3 1 128 36 39 VAL H H 8.343 0.02 1 129 36 39 VAL HA H 3.810 0.02 1 130 36 39 VAL HB H 1.961 0.02 1 131 36 39 VAL N N 121.722 0.3 1 132 37 40 LEU H H 7.039 0.02 1 133 37 40 LEU HA H 3.634 0.02 1 134 37 40 LEU HB2 H 1.668 0.02 2 135 37 40 LEU HB3 H 1.668 0.02 2 136 37 40 LEU N N 120.454 0.3 1 137 38 41 CYS H H 8.200 0.02 1 138 38 41 CYS HA H 4.233 0.02 1 139 38 41 CYS HB2 H 3.169 0.02 2 140 38 41 CYS HB3 H 3.169 0.02 2 141 38 41 CYS N N 117.770 0.3 1 142 40 43 GLU H H 8.839 0.02 1 143 40 43 GLU HA H 4.092 0.02 1 144 40 43 GLU HB2 H 2.192 0.02 2 145 40 43 GLU HB3 H 2.192 0.02 2 146 40 43 GLU HG2 H 2.625 0.02 2 147 40 43 GLU HG3 H 2.625 0.02 2 148 40 43 GLU N N 120.138 0.3 1 149 41 44 ASN H H 8.557 0.02 1 150 41 44 ASN HA H 4.630 0.02 1 151 41 44 ASN HB2 H 3.005 0.02 2 152 41 44 ASN HB3 H 3.005 0.02 2 153 41 44 ASN N N 118.592 0.3 1 154 42 45 ARG H H 7.779 0.02 1 155 42 45 ARG HA H 4.145 0.02 1 156 42 45 ARG HB2 H 1.956 0.02 2 157 42 45 ARG HB3 H 1.956 0.02 2 158 42 45 ARG N N 118.979 0.3 1 159 43 46 LYS H H 7.960 0.02 1 160 43 46 LYS HA H 4.119 0.02 1 161 43 46 LYS HB2 H 1.851 0.02 2 162 43 46 LYS HB3 H 1.851 0.02 2 163 43 46 LYS N N 118.929 0.3 1 164 44 47 ARG H H 8.181 0.02 1 165 44 47 ARG HA H 4.368 0.02 1 166 44 47 ARG HB2 H 1.943 0.02 2 167 44 47 ARG HB3 H 1.943 0.02 2 168 44 47 ARG N N 115.840 0.3 1 169 45 48 GLN H H 8.031 0.02 1 170 45 48 GLN HA H 4.106 0.02 1 171 45 48 GLN HB2 H 2.323 0.02 2 172 45 48 GLN HB3 H 2.323 0.02 2 173 45 48 GLN N N 117.004 0.3 1 174 46 49 THR H H 8.157 0.02 1 175 46 49 THR HA H 4.564 0.02 1 176 46 49 THR HB H 4.253 0.02 1 177 46 49 THR HG2 H 1.110 0.02 1 178 46 49 THR N N 110.056 0.3 1 179 47 50 SER H H 8.639 0.02 1 180 47 50 SER HA H 4.420 0.02 1 181 47 50 SER HB2 H 3.699 0.02 2 182 47 50 SER HB3 H 3.699 0.02 2 183 47 50 SER N N 117.578 0.3 1 184 48 51 ILE H H 7.950 0.02 1 185 48 51 ILE HA H 3.739 0.02 1 186 48 51 ILE HB H 1.983 0.02 1 187 48 51 ILE N N 123.055 0.3 1 188 49 52 LEU H H 7.619 0.02 1 189 49 52 LEU HA H 4.590 0.02 1 190 49 52 LEU HB2 H 1.629 0.02 1 191 49 52 LEU HB3 H 1.262 0.02 1 192 49 52 LEU N N 127.714 0.3 1 193 50 53 ILE H H 8.931 0.02 1 194 50 53 ILE HA H 3.961 0.02 1 195 50 53 ILE HB H 1.747 0.02 1 196 50 53 ILE N N 118.829 0.3 1 197 51 54 GLN H H 9.670 0.02 1 198 51 54 GLN HA H 4.276 0.02 1 199 51 54 GLN HB2 H 1.841 0.02 2 200 51 54 GLN HB3 H 1.841 0.02 2 201 51 54 GLN HG2 H 2.292 0.02 2 202 51 54 GLN HG3 H 2.292 0.02 2 203 51 54 GLN N N 130.524 0.3 1 204 52 55 LYS H H 7.623 0.02 1 205 52 55 LYS HA H 4.600 0.02 1 206 52 55 LYS HB2 H 2.153 0.02 2 207 52 55 LYS HB3 H 2.153 0.02 2 208 52 55 LYS N N 111.862 0.3 1 209 53 56 SER H H 8.625 0.02 1 210 53 56 SER HA H 4.512 0.02 1 211 53 56 SER HB2 H 3.935 0.02 2 212 53 56 SER HB3 H 3.935 0.02 2 213 53 56 SER N N 116.412 0.3 1 214 54 57 GLY H H 8.217 0.02 1 215 54 57 GLY HA2 H 4.253 0.02 1 216 54 57 GLY HA3 H 3.594 0.02 1 217 54 57 GLY N N 113.235 0.3 1 218 56 59 CYS H H 8.140 0.02 1 219 56 59 CYS HA H 4.276 0.02 1 220 56 59 CYS HB2 H 3.136 0.02 1 221 56 59 CYS HB3 H 2.559 0.02 1 222 56 59 CYS N N 122.590 0.3 1 stop_ save_