data_27530 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; apoCaM bound to Cav1.2 IQ ; _BMRB_accession_number 27530 _BMRB_flat_file_name bmr27530.str _Entry_type original _Submission_date 2018-06-28 _Accession_date 2018-06-28 _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 Turner Matthew L. . 2 Ames James B. . 3 Anderson David E. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 115 "15N chemical shifts" 115 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2019-09-17 original BMRB . stop_ _Original_release_date 2018-06-29 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; ApoCaM bound to Cav1.2 IQ ; _Citation_status 'in preparation' _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Turner Matthew L. . 2 Ames James B. . 3 Anderson David E. . stop_ _Journal_abbreviation Cell _Journal_volume . _Journal_issue . _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first . _Page_last . _Year . _Details . loop_ _Keyword Hippocampus 'Voltage Gated' calmodulin cav1.2 stop_ save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name apoCaM _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label apoCaM $apoCaM 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_apoCaM _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common apoCaM _Molecular_mass . _Mol_thiol_state 'all free' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 149 _Mol_residue_sequence ; MADQLTEEQIAEFKEAFSLF DKDGDGTITTKELGTVMRSL GQNPTEAELQDMINEVDADG NGTIDFPEFLTMMARKMKDT DSEEEIREAFRVFDKDGNGY ISAAELRHVMTNLGEKLTDE EVDEMIREADIDGDGQVNYE EFVQMMTAK ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 0 MET 2 1 ALA 3 2 ASP 4 3 GLN 5 4 LEU 6 5 THR 7 6 GLU 8 7 GLU 9 8 GLN 10 9 ILE 11 10 ALA 12 11 GLU 13 12 PHE 14 13 LYS 15 14 GLU 16 15 ALA 17 16 PHE 18 17 SER 19 18 LEU 20 19 PHE 21 20 ASP 22 21 LYS 23 22 ASP 24 23 GLY 25 24 ASP 26 25 GLY 27 26 THR 28 27 ILE 29 28 THR 30 29 THR 31 30 LYS 32 31 GLU 33 32 LEU 34 33 GLY 35 34 THR 36 35 VAL 37 36 MET 38 37 ARG 39 38 SER 40 39 LEU 41 40 GLY 42 41 GLN 43 42 ASN 44 43 PRO 45 44 THR 46 45 GLU 47 46 ALA 48 47 GLU 49 48 LEU 50 49 GLN 51 50 ASP 52 51 MET 53 52 ILE 54 53 ASN 55 54 GLU 56 55 VAL 57 56 ASP 58 57 ALA 59 58 ASP 60 59 GLY 61 60 ASN 62 61 GLY 63 62 THR 64 63 ILE 65 64 ASP 66 65 PHE 67 66 PRO 68 67 GLU 69 68 PHE 70 69 LEU 71 70 THR 72 71 MET 73 72 MET 74 73 ALA 75 74 ARG 76 75 LYS 77 76 MET 78 77 LYS 79 78 ASP 80 79 THR 81 80 ASP 82 81 SER 83 82 GLU 84 83 GLU 85 84 GLU 86 85 ILE 87 86 ARG 88 87 GLU 89 88 ALA 90 89 PHE 91 90 ARG 92 91 VAL 93 92 PHE 94 93 ASP 95 94 LYS 96 95 ASP 97 96 GLY 98 97 ASN 99 98 GLY 100 99 TYR 101 100 ILE 102 101 SER 103 102 ALA 104 103 ALA 105 104 GLU 106 105 LEU 107 106 ARG 108 107 HIS 109 108 VAL 110 109 MET 111 110 THR 112 111 ASN 113 112 LEU 114 113 GLY 115 114 GLU 116 115 LYS 117 116 LEU 118 117 THR 119 118 ASP 120 119 GLU 121 120 GLU 122 121 VAL 123 122 ASP 124 123 GLU 125 124 MET 126 125 ILE 127 126 ARG 128 127 GLU 129 128 ALA 130 129 ASP 131 130 ILE 132 131 ASP 133 132 GLY 134 133 ASP 135 134 GLY 136 135 GLN 137 136 VAL 138 137 ASN 139 138 TYR 140 139 GLU 141 140 GLU 142 141 PHE 143 142 VAL 144 143 GLN 145 144 MET 146 145 MET 147 146 THR 148 147 ALA 149 148 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 $apoCaM 'African clawed frog' 8355 Eukaryota Metazoa Xenopus laevis 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 $apoCaM 'recombinant technology' . Escherichia coli . 'pET 3b' 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 $apoCaM 350 uM '[U-100% 15N]' $apoCaM 350 uM '[U-100% 13C; U-100% 15N]' stop_ save_ ############################ # Computer software used # ############################ save_SPARKY _Saveframe_category software _Name SPARKY _Version . loop_ _Vendor _Address _Electronic_address Goddard . . stop_ loop_ _Task 'chemical shift assignment' stop_ _Details . save_ 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_ ######################### # 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 $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' 0.150 . M pH 7.3 . pH pressure 1 . atm temperature 303 . 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 na indirect . . . 0.251449530 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' 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 apoCaM _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 2 3 ASP H H 8.761 0.031 . 2 2 3 ASP N N 120.729 0.612 . 3 4 5 LEU H H 8.372 0.035 . 4 4 5 LEU N N 122.385 0.595 . 5 5 6 THR H H 8.823 0.032 . 6 5 6 THR N N 113.543 0.611 . 7 6 7 GLU H H 9.084 0.033 . 8 6 7 GLU N N 120.684 0.630 . 9 7 8 GLU H H 8.774 0.036 . 10 7 8 GLU N N 119.806 0.601 . 11 8 9 GLN H H 7.795 0.026 . 12 8 9 GLN N N 120.902 0.658 . 13 9 10 ILE H H 8.313 0.036 . 14 9 10 ILE N N 119.267 0.200 . 15 10 11 ALA H H 7.987 0.030 . 16 10 11 ALA N N 120.953 0.580 . 17 11 12 GLU H H 7.882 0.027 . 18 11 12 GLU N N 120.585 0.606 . 19 12 13 PHE H H 8.927 0.033 . 20 12 13 PHE N N 120.968 0.656 . 21 13 14 LYS H H 9.269 0.034 . 22 13 14 LYS N N 122.413 0.344 . 23 14 15 GLU H H 8.121 0.029 . 24 14 15 GLU N N 120.915 0.570 . 25 15 16 ALA H H 7.736 0.031 . 26 15 16 ALA N N 121.397 0.598 . 27 16 17 PHE H H 8.631 0.031 . 28 16 17 PHE N N 118.434 0.571 . 29 17 18 SER H H 8.478 0.032 . 30 17 18 SER N N 111.550 0.555 . 31 18 19 LEU H H 7.424 0.034 . 32 18 19 LEU N N 121.668 0.559 . 33 19 20 PHE H H 7.475 0.051 . 34 19 20 PHE N N 115.463 0.306 . 35 20 21 ASP H H 7.903 0.037 . 36 20 21 ASP N N 122.163 0.338 . 37 21 22 LYS H H 8.451 0.034 . 38 21 22 LYS N N 125.278 0.378 . 39 22 23 ASP H H 8.677 0.036 . 40 22 23 ASP N N 116.888 0.565 . 41 23 24 GLY H H 8.127 0.038 . 42 23 24 GLY N N 110.718 0.407 . 43 24 25 ASP H H 8.825 0.028 . 44 24 25 ASP N N 120.867 0.679 . 45 25 26 GLY H H 10.301 0.129 . 46 25 26 GLY N N 112.520 0.459 . 47 27 28 ILE H H 8.355 0.034 . 48 27 28 ILE N N 111.220 0.281 . 49 28 29 THR H H 8.399 0.031 . 50 28 29 THR N N 111.215 0.605 . 51 29 30 THR H H 8.379 0.032 . 52 29 30 THR N N 112.995 0.562 . 53 30 31 LYS H H 7.685 0.030 . 54 30 31 LYS N N 119.580 0.375 . 55 31 32 GLU H H 7.592 0.031 . 56 31 32 GLU N N 117.906 0.765 . 57 32 33 LEU H H 7.458 0.035 . 58 32 33 LEU N N 121.671 0.311 . 59 33 34 GLY H H 8.871 0.010 . 60 33 34 GLY N N 105.991 0.632 . 61 34 35 THR H H 7.587 0.032 . 62 34 35 THR N N 118.924 0.565 . 63 35 36 VAL H H 8.042 0.031 . 64 35 36 VAL N N 123.253 0.588 . 65 36 37 MET H H 8.530 0.029 . 66 36 37 MET N N 119.122 0.598 . 67 37 38 ARG H H 8.584 0.031 . 68 37 38 ARG N N 120.006 0.557 . 69 38 39 SER H H 8.188 0.025 . 70 38 39 SER N N 119.606 0.573 . 71 39 40 LEU H H 7.436 0.036 . 72 39 40 LEU N N 121.893 0.333 . 73 40 41 GLY H H 7.986 0.032 . 74 40 41 GLY N N 107.650 0.614 . 75 42 43 ASN H H 8.736 0.031 . 76 42 43 ASN N N 117.092 0.595 . 77 45 46 GLU H H 8.949 0.055 . 78 45 46 GLU N N 121.046 0.535 . 79 46 47 ALA H H 8.391 0.034 . 80 46 47 ALA N N 121.908 0.333 . 81 47 48 GLU H H 7.836 0.039 . 82 47 48 GLU N N 119.590 0.590 . 83 48 49 LEU H H 8.400 0.036 . 84 48 49 LEU N N 120.998 0.286 . 85 50 51 ASP H H 7.941 0.029 . 86 50 51 ASP N N 119.557 0.676 . 87 52 53 ILE H H 8.400 0.036 . 88 52 53 ILE N N 120.017 0.285 . 89 53 54 ASN H H 8.354 0.030 . 90 53 54 ASN N N 117.982 0.592 . 91 54 55 GLU H H 7.681 0.035 . 92 54 55 GLU N N 117.806 0.644 . 93 56 57 ASP H H 8.504 0.025 . 94 56 57 ASP N N 122.362 0.560 . 95 57 58 ALA H H 8.205 0.030 . 96 57 58 ALA N N 125.496 0.640 . 97 58 59 ASP H H 8.482 0.030 . 98 58 59 ASP N N 115.521 0.621 . 99 59 60 GLY H H 7.994 0.033 . 100 59 60 GLY N N 109.393 0.581 . 101 60 61 ASN H H 9.245 0.029 . 102 60 61 ASN N N 120.090 0.603 . 103 61 62 GLY H H 9.921 0.031 . 104 61 62 GLY N N 110.447 0.592 . 105 63 64 ILE H H 8.996 0.036 . 106 63 64 ILE N N 119.571 0.645 . 107 64 65 ASP H H 8.364 0.033 . 108 64 65 ASP N N 124.499 0.594 . 109 65 66 PHE H H 8.751 0.032 . 110 65 66 PHE N N 119.191 0.450 . 111 67 68 GLU H H 8.177 0.032 . 112 67 68 GLU N N 118.055 0.509 . 113 68 69 PHE H H 8.573 0.035 . 114 68 69 PHE N N 123.010 0.594 . 115 69 70 LEU H H 8.538 0.030 . 116 69 70 LEU N N 119.108 0.581 . 117 72 73 MET H H 8.131 0.041 . 118 72 73 MET N N 118.022 0.606 . 119 73 74 ALA H H 8.366 0.043 . 120 73 74 ALA N N 121.268 0.624 . 121 74 75 ARG H H 7.558 0.035 . 122 74 75 ARG N N 117.098 0.931 . 123 75 76 LYS H H 7.791 0.020 . 124 75 76 LYS N N 119.015 0.689 . 125 77 78 LYS H H 7.839 0.040 . 126 77 78 LYS N N 121.088 0.616 . 127 78 79 ASP H H 8.315 0.015 . 128 78 79 ASP N N 122.386 0.448 . 129 80 81 ASP H H 8.519 0.033 . 130 80 81 ASP N N 123.604 0.588 . 131 81 82 SER H H 8.534 0.037 . 132 81 82 SER N N 117.619 0.599 . 133 82 83 GLU H H 8.472 0.036 . 134 82 83 GLU N N 122.849 0.652 . 135 83 84 GLU H H 8.235 0.031 . 136 83 84 GLU N N 119.328 0.216 . 137 84 85 GLU H H 8.092 0.026 . 138 84 85 GLU N N 119.697 0.508 . 139 85 86 ILE H H 8.004 0.036 . 140 85 86 ILE N N 120.838 0.286 . 141 87 88 GLU H H 8.383 0.019 . 142 87 88 GLU N N 118.228 0.996 . 143 88 89 ALA H H 7.597 0.031 . 144 88 89 ALA N N 121.819 0.665 . 145 89 90 PHE H H 7.570 0.044 . 146 89 90 PHE N N 115.003 0.300 . 147 91 92 VAL H H 7.221 0.032 . 148 91 92 VAL N N 117.400 0.705 . 149 92 93 PHE H H 7.424 0.031 . 150 92 93 PHE N N 116.338 0.731 . 151 93 94 ASP H H 7.384 0.033 . 152 93 94 ASP N N 122.885 0.607 . 153 94 95 LYS H H 8.134 0.029 . 154 94 95 LYS N N 124.127 0.581 . 155 95 96 ASP H H 8.787 0.034 . 156 95 96 ASP N N 117.493 0.558 . 157 96 97 GLY H H 8.083 0.032 . 158 96 97 GLY N N 110.756 0.628 . 159 98 99 GLY H H 10.085 0.095 . 160 98 99 GLY N N 111.119 0.300 . 161 100 101 ILE H H 8.769 0.058 . 162 100 101 ILE N N 113.579 0.626 . 163 101 102 SER H H 9.023 0.076 . 164 101 102 SER N N 118.580 0.655 . 165 102 103 ALA H H 8.935 0.052 . 166 102 103 ALA N N 125.455 0.733 . 167 103 104 ALA H H 8.352 0.044 . 168 103 104 ALA N N 119.788 0.274 . 169 104 105 GLU H H 7.812 0.034 . 170 104 105 GLU N N 120.522 0.197 . 171 105 106 LEU H H 8.272 0.028 . 172 105 106 LEU N N 121.728 0.524 . 173 107 108 HIS H H 7.789 0.028 . 174 107 108 HIS N N 118.815 0.558 . 175 108 109 VAL H H 8.204 0.040 . 176 108 109 VAL N N 120.586 0.276 . 177 110 111 THR H H 8.047 0.062 . 178 110 111 THR N N 112.764 0.988 . 179 111 112 ASN H H 7.783 0.016 . 180 111 112 ASN N N 120.725 0.675 . 181 113 114 GLY H H 8.223 0.037 . 182 113 114 GLY N N 108.896 0.557 . 183 114 115 GLU H H 8.202 0.051 . 184 114 115 GLU N N 120.619 0.240 . 185 115 116 LYS H H 8.393 0.029 . 186 115 116 LYS N N 121.552 0.438 . 187 116 117 LEU H H 7.828 0.033 . 188 116 117 LEU N N 121.874 0.504 . 189 117 118 THR H H 9.007 0.034 . 190 117 118 THR N N 113.917 0.610 . 191 118 119 ASP H H 8.886 0.034 . 192 118 119 ASP N N 121.427 0.648 . 193 120 121 GLU H H 7.893 0.043 . 194 120 121 GLU N N 121.312 0.548 . 195 121 122 VAL H H 8.304 0.021 . 196 121 122 VAL N N 121.427 0.646 . 197 122 123 ASP H H 8.445 0.030 . 198 122 123 ASP N N 119.830 0.649 . 199 124 125 MET H H 8.025 0.038 . 200 124 125 MET N N 119.523 0.650 . 201 125 126 ILE H H 8.439 0.034 . 202 125 126 ILE N N 119.412 0.229 . 203 126 127 ARG H H 8.011 0.034 . 204 126 127 ARG N N 119.903 0.568 . 205 127 128 GLU H H 7.928 0.049 . 206 127 128 GLU N N 117.216 0.654 . 207 128 129 ALA H H 7.665 0.036 . 208 128 129 ALA N N 121.585 0.608 . 209 129 130 ASP H H 8.358 0.031 . 210 129 130 ASP N N 120.662 0.399 . 211 131 132 ASP H H 8.671 0.036 . 212 131 132 ASP N N 124.246 0.528 . 213 132 133 GLY H H 8.412 0.033 . 214 132 133 GLY N N 108.497 0.609 . 215 133 134 ASP H H 8.364 0.030 . 216 133 134 ASP N N 120.177 0.215 . 217 134 135 GLY H H 8.571 0.094 . 218 134 135 GLY N N 109.772 0.806 . 219 136 137 VAL H H 9.441 0.043 . 220 136 137 VAL N N 120.172 0.300 . 221 138 139 TYR H H 7.750 0.089 . 222 138 139 TYR N N 123.067 0.789 . 223 139 140 GLU H H 8.204 0.046 . 224 139 140 GLU N N 126.592 0.375 . 225 145 146 MET H H 8.140 0.077 . 226 145 146 MET N N 116.091 0.886 . 227 147 148 ALA H H 7.676 0.038 . 228 147 148 ALA N N 126.405 0.486 . 229 148 149 LYS H H 7.848 0.085 . 230 148 149 LYS N N 126.504 0.460 . stop_ save_