data_25303 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Titin M10 bound to obscurin ig1 ; _BMRB_accession_number 25303 _BMRB_flat_file_name bmr25303.str _Entry_type original _Submission_date 2014-10-29 _Accession_date 2014-10-29 _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 Wright Nathan T. . 2 Rudloff Michael W. . 3 Woosley Alec N. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 79 "15N chemical shifts" 79 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2015-08-25 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 25301 OBSC-Ig1 25304 'obscurin Ig1 bound to titin M10' 25305 'titin M10' 25308 'obscurin Ig58' 25406 'H56P mutant' stop_ _Original_release_date 2015-08-25 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Biophysical characterization of naturally occurring titin M10 mutations ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 25739468 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Rudloff Michael W. . 2 Woosley Alec N. . 3 Wright Nathan T. . stop_ _Journal_abbreviation 'Protein Sci.' _Journal_volume 24 _Journal_issue 6 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 946 _Page_last 955 _Year 2015 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'titin M10-obscurin-Ig1' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'titin M10' $titin_M10 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_titin_M10 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common titin_M10 _Molecular_mass . _Mol_thiol_state unknown _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 101 _Mol_residue_sequence ; GSRGIPPKIEALPSDISIDE GKVLTVACAFTGEPTPEVTW SCGGRKIHSQEQGRFHIENT DDLTTLIIMDVQKQDGGLYT LSLGNEFGSDSATVNIHIRS I ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 1 GLY 2 2 SER 3 3 ARG 4 4 GLY 5 5 ILE 6 6 PRO 7 7 PRO 8 8 LYS 9 9 ILE 10 10 GLU 11 11 ALA 12 12 LEU 13 13 PRO 14 14 SER 15 15 ASP 16 16 ILE 17 17 SER 18 18 ILE 19 19 ASP 20 20 GLU 21 21 GLY 22 22 LYS 23 23 VAL 24 24 LEU 25 25 THR 26 26 VAL 27 27 ALA 28 28 CYS 29 29 ALA 30 30 PHE 31 31 THR 32 32 GLY 33 33 GLU 34 34 PRO 35 35 THR 36 36 PRO 37 37 GLU 38 38 VAL 39 39 THR 40 40 TRP 41 41 SER 42 42 CYS 43 43 GLY 44 44 GLY 45 45 ARG 46 46 LYS 47 47 ILE 48 48 HIS 49 49 SER 50 50 GLN 51 51 GLU 52 52 GLN 53 53 GLY 54 54 ARG 55 55 PHE 56 56 HIS 57 57 ILE 58 58 GLU 59 59 ASN 60 60 THR 61 61 ASP 62 62 ASP 63 63 LEU 64 64 THR 65 65 THR 66 66 LEU 67 67 ILE 68 68 ILE 69 69 MET 70 70 ASP 71 71 VAL 72 72 GLN 73 73 LYS 74 74 GLN 75 75 ASP 76 76 GLY 77 77 GLY 78 78 LEU 79 79 TYR 80 80 THR 81 81 LEU 82 82 SER 83 83 LEU 84 84 GLY 85 85 ASN 86 86 GLU 87 87 PHE 88 88 GLY 89 89 SER 90 90 ASP 91 91 SER 92 92 ALA 93 93 THR 94 94 VAL 95 95 ASN 96 96 ILE 97 97 HIS 98 98 ILE 99 99 ARG 100 100 SER 101 101 ILE stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-11-25 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value BMRB 25305 titin_M10 100.00 101 100.00 100.00 1.33e-66 BMRB 25406 H56P 100.00 107 99.01 99.01 8.94e-65 PDB 2WP3 "Crystal Structure Of The Titin M10-Obscurin Like 1 Ig Complex" 99.01 102 100.00 100.00 6.24e-66 PDB 2WWK "Crystal Structure Of The Titin M10-Obscurin Like 1 Ig F17r Mutant Complex" 99.01 102 100.00 100.00 6.24e-66 PDB 2WWM "Crystal Structure Of The Titin M10-Obscurin Like 1 Ig Complex In Space Group P1" 99.01 102 100.00 100.00 6.24e-66 PDB 2Y9R "Crystal Structure Of The M10 Domain Of Titin" 99.01 102 100.00 100.00 6.24e-66 PDB 3KNB "Crystal Structure Of The Titin C-Terminus In Complex With Obscurin- Like 1" 97.03 100 100.00 100.00 4.04e-64 PDB 3Q5O "Crystal Structure Of Human Titin Domain M10" 97.03 100 100.00 100.00 4.04e-64 PDB 4C4K "Crystal Structure Of The Titin M10-obscurin Ig Domain 1 Complex" 99.01 102 100.00 100.00 6.24e-66 PDB 4UOW "Crystal Structure Of The Titin M10-obscurin Ig Domain 1 Complex" 96.04 97 100.00 100.00 8.83e-64 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $titin_M10 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 $titin_M10 'recombinant technology' . Escherichia coli . pET24a stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details 'All NMR samples were collected at 25oC in 20 mM Tris pH 7.5, 20 mM NaCl, 0.35 mM NaN3, and 0.5-2.5 mM protein with 10% D2O.' loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $titin_M10 . mM 0.5 2.5 '[U-99% 13C; U-99% 15N]' Tris 20 mM . . 'natural abundance' NaCl 20 mM . . 'natural abundance' NaN3 0.35 mM . . 'natural abundance' H2O 90 % . . 'natural abundance' D2O 10 % . . 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ 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_ save_3D_CBCA(CO)NH_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D CBCA(CO)NH' _Sample_label $sample_1 save_ save_3D_HNCO_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCO' _Sample_label $sample_1 save_ save_3D_HCACO_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HCACO' _Sample_label $sample_1 save_ save_3D_HNCACB_5 _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 'All NMR samples were collected at 25oC in 20 mM Tris pH 7.5, 20 mM NaCl, 0.35 mM NaN3, and 0.5-2.5 mM protein with 10% D2O.' loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' .02 . M pH 7.5 . 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 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' '3D CBCA(CO)NH' '3D HNCO' '3D HCACO' '3D HNCACB' 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 'titin M10' _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 4 4 GLY H H 7.386 0.002 1 2 4 4 GLY N N 109.92 0.01 1 3 5 5 ILE H H 9.130 0.002 1 4 5 5 ILE N N 121.81 0.01 1 5 10 10 GLU H H 8.753 0.002 1 6 10 10 GLU N N 129.72 0.01 1 7 11 11 ALA H H 6.818 0.002 1 8 11 11 ALA N N 114.87 0.01 1 9 12 12 LEU H H 8.689 0.002 1 10 12 12 LEU N N 127.02 0.01 1 11 14 14 SER H H 8.405 0.002 1 12 14 14 SER N N 113.73 0.01 1 13 15 15 ASP H H 7.510 0.002 1 14 15 15 ASP N N 120.67 0.01 1 15 16 16 ILE H H 8.759 0.002 1 16 16 16 ILE N N 121.09 0.01 1 17 17 17 SER H H 7.806 0.002 1 18 17 17 SER N N 119.56 0.01 1 19 19 19 ASP H H 7.909 0.002 1 20 19 19 ASP N N 125.52 0.01 1 21 20 20 GLU H H 8.013 0.002 1 22 20 20 GLU N N 118.24 0.01 1 23 21 21 GLY H H 9.812 0.002 1 24 21 21 GLY N N 112.52 0.01 1 25 22 22 LYS H H 7.174 0.002 1 26 22 22 LYS N N 119.29 0.01 1 27 25 25 THR H H 8.625 0.002 1 28 25 25 THR N N 120.16 0.01 1 29 26 26 VAL H H 9.388 0.002 1 30 26 26 VAL N N 126.37 0.01 1 31 27 27 ALA H H 8.556 0.002 1 32 27 27 ALA N N 128.71 0.01 1 33 28 28 CYS H H 9.517 0.002 1 34 28 28 CYS N N 121.53 0.01 1 35 29 29 ALA H H 9.470 0.002 1 36 29 29 ALA N N 133.16 0.01 1 37 30 30 PHE H H 7.551 0.002 1 38 30 30 PHE N N 114.50 0.01 1 39 31 31 THR H H 8.478 0.002 1 40 31 31 THR N N 110.17 0.01 1 41 32 32 GLY H H 8.356 0.002 1 42 32 32 GLY N N 107.40 0.01 1 43 35 35 THR H H 8.581 0.002 1 44 35 35 THR N N 118.82 0.01 1 45 37 37 GLU H H 8.747 0.002 1 46 37 37 GLU N N 119.88 0.01 1 47 38 38 VAL H H 8.596 0.002 1 48 38 38 VAL N N 129.78 0.01 1 49 40 40 TRP H H 9.658 0.002 1 50 40 40 TRP N N 130.10 0.01 1 51 41 41 SER H H 9.415 0.002 1 52 41 41 SER N N 115.04 0.01 1 53 42 42 CYS H H 8.534 0.002 1 54 42 42 CYS N N 120.42 0.01 1 55 45 45 ARG H H 7.616 0.002 1 56 45 45 ARG N N 121.15 0.01 1 57 46 46 LYS H H 8.415 0.002 1 58 46 46 LYS N N 126.28 0.01 1 59 47 47 ILE H H 8.806 0.002 1 60 47 47 ILE N N 126.66 0.01 1 61 48 48 HIS H H 8.232 0.002 1 62 48 48 HIS N N 125.47 0.01 1 63 52 52 GLN H H 7.985 0.002 1 64 52 52 GLN N N 119.57 0.01 1 65 53 53 GLY H H 8.115 0.002 1 66 53 53 GLY N N 108.32 0.01 1 67 55 55 PHE H H 7.738 0.002 1 68 55 55 PHE N N 117.34 0.01 1 69 56 56 HIS H H 9.074 0.002 1 70 56 56 HIS N N 124.68 0.01 1 71 57 57 ILE H H 8.162 0.002 1 72 57 57 ILE N N 122.52 0.01 1 73 58 58 GLU H H 9.012 0.002 1 74 58 58 GLU N N 128.14 0.01 1 75 59 59 ASN H H 9.232 0.002 1 76 59 59 ASN N N 126.28 0.01 1 77 60 60 THR H H 9.312 0.002 1 78 60 60 THR N N 117.12 0.01 1 79 61 61 ASP H H 8.377 0.002 1 80 61 61 ASP N N 117.45 0.01 1 81 62 62 ASP H H 7.962 0.002 1 82 62 62 ASP N N 115.87 0.01 1 83 63 63 LEU H H 7.067 0.002 1 84 63 63 LEU N N 121.18 0.01 1 85 65 65 THR H H 8.574 0.002 1 86 65 65 THR N N 125.28 0.01 1 87 66 66 LEU H H 8.858 0.002 1 88 66 66 LEU N N 129.77 0.01 1 89 67 67 ILE H H 8.998 0.002 1 90 67 67 ILE N N 127.31 0.01 1 91 68 68 ILE H H 8.848 0.002 1 92 68 68 ILE N N 123.35 0.01 1 93 69 69 MET H H 8.276 0.002 1 94 69 69 MET N N 122.61 0.01 1 95 70 70 ASP H H 7.001 0.002 1 96 70 70 ASP N N 118.93 0.01 1 97 71 71 VAL H H 8.129 0.002 1 98 71 71 VAL N N 115.21 0.01 1 99 72 72 GLN H H 9.262 0.002 1 100 72 72 GLN N N 125.42 0.01 1 101 73 73 LYS H H 9.238 0.002 1 102 73 73 LYS N N 122.07 0.01 1 103 74 74 GLN H H 8.810 0.002 1 104 74 74 GLN N N 115.66 0.01 1 105 75 75 ASP H H 8.211 0.002 1 106 75 75 ASP N N 119.96 0.01 1 107 76 76 GLY H H 7.633 0.002 1 108 76 76 GLY N N 105.66 0.01 1 109 77 77 GLY H H 8.869 0.002 1 110 77 77 GLY N N 109.72 0.01 1 111 78 78 LEU H H 8.268 0.002 1 112 78 78 LEU N N 120.55 0.01 1 113 79 79 TYR H H 9.829 0.002 1 114 79 79 TYR N N 131.68 0.01 1 115 80 80 THR H H 9.287 0.002 1 116 80 80 THR N N 118.21 0.01 1 117 81 81 LEU H H 8.860 0.002 1 118 81 81 LEU N N 131.05 0.01 1 119 82 82 SER H H 8.942 0.002 1 120 82 82 SER N N 119.86 0.01 1 121 83 83 LEU H H 8.717 0.002 1 122 83 83 LEU N N 122.07 0.01 1 123 84 84 GLY H H 7.934 0.002 1 124 84 84 GLY N N 106.38 0.01 1 125 85 85 ASN H H 8.959 0.002 1 126 85 85 ASN N N 121.30 0.01 1 127 86 86 GLU H H 9.209 0.002 1 128 86 86 GLU N N 116.32 0.01 1 129 87 87 PHE H H 7.923 0.002 1 130 87 87 PHE N N 115.85 0.01 1 131 88 88 GLY H H 7.760 0.002 1 132 88 88 GLY N N 107.28 0.01 1 133 89 89 SER H H 8.458 0.002 1 134 89 89 SER N N 112.41 0.01 1 135 90 90 ASP H H 8.468 0.002 1 136 90 90 ASP N N 120.54 0.01 1 137 91 91 SER H H 8.202 0.002 1 138 91 91 SER N N 115.49 0.01 1 139 92 92 ALA H H 8.809 0.002 1 140 92 92 ALA N N 124.13 0.01 1 141 93 93 THR H H 8.305 0.002 1 142 93 93 THR N N 111.35 0.01 1 143 94 94 VAL H H 9.438 0.002 1 144 94 94 VAL N N 122.42 0.01 1 145 95 95 ASN H H 8.423 0.002 1 146 95 95 ASN N N 128.26 0.01 1 147 96 96 ILE H H 9.004 0.002 1 148 96 96 ILE N N 126.56 0.01 1 149 97 97 HIS H H 9.205 0.002 1 150 97 97 HIS N N 129.20 0.01 1 151 98 98 ILE H H 8.547 0.002 1 152 98 98 ILE N N 124.31 0.01 1 153 99 99 ARG H H 8.882 0.002 1 154 99 99 ARG N N 128.96 0.01 1 155 100 100 SER H H 8.440 0.002 1 156 100 100 SER N N 120.02 0.01 1 157 101 101 ILE H H 8.187 0.002 1 158 101 101 ILE N N 127.12 0.01 1 stop_ save_