data_6260 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; NMR solution Structure of a Highly Stable de novo Heterodimeric Coiled-Coil ; _BMRB_accession_number 6260 _BMRB_flat_file_name bmr6260.str _Entry_type original _Submission_date 2004-07-12 _Accession_date 2004-07-12 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details ; Chemical shifts for E3/K3 coiled-coil are deposited as a single file, with a non-existent poly-glycine linker connecting the two subunits, as required for calculations using CYANA. Please disregard the Poly-Gly in the reported sequence. ; loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Lindhout Darrin A. . 2 Litowski Jennifer R. . 3 Mercier Pascal . . 4 Hodges Robert S. . 5 Sykes Brian D. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 2 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 186 "15N chemical shifts" 42 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2004-07-26 original author . stop_ _Original_release_date 2004-07-26 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'NMR solution Structure of a Highly Stable de novo Heterodimeric Coiled-Coil' _Citation_status submitted _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 Lindhout Darrin A. . 2 Litowski Jennifer R. . 3 Mercier Pascal . . 4 Hodges Robert S. . 5 Sykes Brian D. . stop_ _Journal_abbreviation Biopolymers _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 IAAL-E3/K3 stop_ save_ ################################## # Molecular system description # ################################## save_system_EK_coiled-coil _Saveframe_category molecular_system _Mol_system_name 'IAAL-E3/K3 heterodimer' _Abbreviation_common 'EK coiled-coil' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label IAAL-E3 $E3 'K3 coiled-coil' $K3 stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state heterodimer _System_paramagnetic no _System_thiol_state 'not present' _Database_query_date . _Details ; During CYANA structural calculations, a poly-gly linker was introduced. The sequence thus contains a non-existent gly-linker (total residues = 51), when in fact there were two seperate synthetic peptides constructed, each of 21 residues. The poly-gly is a remnant of structure calculations: EIAALEKEIAALEKEIAALEKGGGGGGGGGKIAALKEKIAALKEKIAALKE ; save_ ######################## # Monomeric polymers # ######################## save_E3 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common IAAL-E3 _Abbreviation_common IAAL-E3 _Molecular_mass . _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 21 _Mol_residue_sequence ; EIAALEKEIAALEKEIAALE K ; loop_ _Residue_seq_code _Residue_label 1 GLU 2 ILE 3 ALA 4 ALA 5 LEU 6 GLU 7 LYS 8 GLU 9 ILE 10 ALA 11 ALA 12 LEU 13 GLU 14 LYS 15 GLU 16 ILE 17 ALA 18 ALA 19 LEU 20 GLU 21 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2014-10-26 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 PDB 1U0I "Iaal-E3K3 HETERODIMER" 100.00 21 100.00 100.00 2.66e-01 PDB 4ETP "C-terminal Motor And Motor Homology Domain Of Kar3vik1 Fused To A Synthetic Heterodimeric Coiled Coil" 100.00 403 100.00 100.00 4.72e-01 stop_ save_ save_K3 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'K3 coiled-coil' _Abbreviation_common 'K3 coiled-coil' _Molecular_mass . _Mol_thiol_state 'not present' _Details . _Residue_count 21 _Mol_residue_sequence ; KIAALKEKIAALKEKIAALK E ; loop_ _Residue_seq_code _Residue_label 1 LYS 2 ILE 3 ALA 4 ALA 5 LEU 6 LYS 7 GLU 8 LYS 9 ILE 10 ALA 11 ALA 12 LEU 13 LYS 14 GLU 15 LYS 16 ILE 17 ALA 18 ALA 19 LEU 20 LYS 21 GLU stop_ _Sequence_homology_query_date 2008-08-19 _Sequence_homology_query_revised_last_date 2008-08-19 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 PDB 1U0I 'Iaal-E3K3 HETERODIMER' 100.00 21 100.00 100.00 2.64e+00 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Details $E3 . . . . . . 'synthetic peptide, unclassified' $K3 . . . . . . . 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 $E3 'chemical synthesis' . . . . . $K3 'chemical synthesis' . . . . . 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 $E3 1.0 mM . $K3 1.0 mM . stop_ save_ ############################ # Computer software used # ############################ save_vnmr _Saveframe_category software _Name VNMR _Version . _Details . save_ save_cyana _Saveframe_category software _Name CYANA _Version . _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model INOVA _Field_strength 500 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model INOVA _Field_strength 800 _Details . save_ ############################# # NMR applied experiments # ############################# save_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC' _Sample_label . save_ save_1H-1H_NOESY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H NOESY' _Sample_label . save_ save_1H-1H_TOCSY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H TOCSY' _Sample_label . save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_experimental _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 150 . mM pH 6.70 0.02 n/a temperature 293 0.05 K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference _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 0.0 internal direct . . . 1.0 DSS N 15 'methyl protons' ppm 0.0 . 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_shift_set_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $experimental _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name IAAL-E3 _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 GLU N N 127.040 0.05 1 2 . 1 GLU H H 8.525 0.02 1 3 . 1 GLU HA H 4.221 0.02 1 4 . 1 GLU HB3 H 2.092 0.02 2 5 . 1 GLU HB2 H 2.021 0.02 2 6 . 1 GLU HG3 H 2.355 0.02 2 7 . 2 ILE N N 119.971 0.05 1 8 . 2 ILE H H 8.170 0.02 1 9 . 2 ILE HA H 3.953 0.02 1 10 . 2 ILE HB H 1.954 0.02 1 11 . 2 ILE HG12 H 1.301 0.02 1 12 . 2 ILE HD1 H 0.949 0.02 1 13 . 2 ILE HG2 H 0.910 0.02 1 14 . 3 ALA N N 123.043 0.05 1 15 . 3 ALA H H 8.085 0.02 1 16 . 3 ALA HA H 4.260 0.02 1 17 . 3 ALA HB H 1.557 0.02 1 18 . 4 ALA N N 120.598 0.05 1 19 . 4 ALA H H 7.887 0.02 1 20 . 4 ALA HA H 4.187 0.02 1 21 . 4 ALA HB H 1.611 0.02 1 22 . 5 LEU N N 121.626 0.05 1 23 . 5 LEU H H 8.207 0.02 1 24 . 5 LEU HA H 4.184 0.02 1 25 . 5 LEU HB3 H 1.892 0.02 2 26 . 5 LEU HB2 H 1.813 0.02 2 27 . 5 LEU HD1 H 0.924 0.02 2 28 . 5 LEU HD2 H 0.961 0.02 2 29 . 6 GLU N N 118.245 0.05 1 30 . 6 GLU H H 8.786 0.02 1 31 . 6 GLU HA H 3.887 0.02 1 32 . 6 GLU HB3 H 2.071 0.02 2 33 . 6 GLU HB2 H 2.180 0.02 2 34 . 6 GLU HG3 H 2.246 0.02 2 35 . 6 GLU HG2 H 2.620 0.02 2 36 . 7 LYS N N 118.720 0.05 1 37 . 7 LYS H H 7.770 0.02 1 38 . 7 LYS HA H 4.113 0.02 1 39 . 7 LYS HB3 H 2.037 0.02 2 40 . 7 LYS HG2 H 1.514 0.02 2 41 . 7 LYS HD2 H 1.693 0.02 2 42 . 7 LYS HE2 H 3.019 0.02 2 43 . 8 GLU N N 120.581 0.05 1 44 . 8 GLU H H 7.911 0.02 1 45 . 8 GLU HA H 4.184 0.02 1 46 . 8 GLU HB3 H 2.216 0.02 2 47 . 8 GLU HB2 H 2.276 0.02 2 48 . 8 GLU HG3 H 2.411 0.02 2 49 . 9 ILE N N 119.874 0.05 1 50 . 9 ILE H H 8.554 0.02 1 51 . 9 ILE HA H 3.689 0.02 1 52 . 9 ILE HB H 1.945 0.02 1 53 . 9 ILE HG13 H 1.106 0.02 1 54 . 9 ILE HG12 H 0.949 0.02 1 55 . 9 ILE HD1 H 0.922 0.02 1 56 . 9 ILE HG2 H 0.842 0.02 1 57 . 10 ALA N N 121.327 0.05 1 58 . 10 ALA H H 8.086 0.02 1 59 . 10 ALA HA H 4.260 0.02 1 60 . 10 ALA HB H 1.480 0.02 1 61 . 11 ALA N N 118.591 0.05 1 62 . 11 ALA H H 7.858 0.02 1 63 . 11 ALA HA H 4.198 0.02 1 64 . 11 ALA HB H 1.548 0.02 1 65 . 12 LEU N N 120.643 0.05 1 66 . 12 LEU H H 7.954 0.02 1 67 . 12 LEU HA H 4.205 0.02 1 68 . 12 LEU HB3 H 1.806 0.02 2 69 . 13 GLU N N 117.927 0.05 1 70 . 13 GLU H H 8.529 0.02 1 71 . 13 GLU HA H 3.895 0.02 1 72 . 13 GLU HB3 H 2.208 0.02 2 73 . 13 GLU HB2 H 2.075 0.02 2 74 . 13 GLU HG3 H 2.545 0.02 2 75 . 13 GLU HG2 H 2.620 0.02 2 76 . 14 LYS N N 118.630 0.05 1 77 . 14 LYS H H 7.791 0.02 1 78 . 14 LYS HA H 4.121 0.02 1 79 . 14 LYS HE2 H 3.020 0.02 2 80 . 15 GLU N N 121.077 0.05 1 81 . 15 GLU H H 7.945 0.02 1 82 . 15 GLU HA H 4.198 0.02 1 83 . 15 GLU HB3 H 2.203 0.02 2 84 . 15 GLU HB2 H 2.269 0.02 2 85 . 15 GLU HG3 H 2.400 0.02 2 86 . 16 ILE N N 119.874 0.05 1 87 . 16 ILE H H 8.557 0.02 1 88 . 16 ILE HA H 3.687 0.02 1 89 . 16 ILE HB H 1.944 0.02 1 90 . 17 ALA N N 121.424 0.05 1 91 . 17 ALA H H 8.120 0.02 1 92 . 17 ALA HA H 4.181 0.02 1 93 . 17 ALA HB H 1.533 0.02 1 94 . 18 ALA N N 119.388 0.05 1 95 . 18 ALA H H 7.739 0.02 1 96 . 18 ALA HA H 4.186 0.02 1 97 . 18 ALA HB H 1.569 0.02 1 98 . 19 LEU N N 119.903 0.05 1 99 . 19 LEU H H 7.980 0.02 1 100 . 19 LEU HA H 4.182 0.02 1 101 . 19 LEU HB3 H 2.005 0.02 2 102 . 19 LEU HG H 1.641 0.02 1 103 . 19 LEU HD1 H 0.953 0.02 2 104 . 20 GLU N N 118.263 0.05 1 105 . 20 GLU H H 8.260 0.02 1 106 . 20 GLU HA H 4.124 0.02 1 107 . 20 GLU HB3 H 2.121 0.02 2 108 . 20 GLU HB2 H 2.260 0.02 2 109 . 20 GLU HG3 H 2.499 0.02 2 110 . 20 GLU HG2 H 2.520 0.02 2 111 . 21 LYS N N 119.269 0.05 1 112 . 21 LYS H H 7.806 0.02 1 113 . 21 LYS HA H 4.235 0.02 1 114 . 21 LYS HB3 H 1.977 0.02 2 115 . 21 LYS HB2 H 1.929 0.02 2 stop_ save_ save_shift_set_2 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $experimental _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'K3 coiled-coil' _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 LYS N N 127.345 0.05 1 2 . 1 LYS H H 8.370 0.02 1 3 . 1 LYS HA H 4.137 0.02 1 4 . 1 LYS HB3 H 1.863 0.02 2 5 . 1 LYS HB2 H 2.159 0.02 2 6 . 1 LYS HG3 H 1.485 0.02 2 7 . 1 LYS HG2 H 1.593 0.02 2 8 . 1 LYS HD2 H 1.758 0.02 2 9 . 1 LYS HE2 H 3.014 0.02 2 10 . 2 ILE N N 118.369 0.05 1 11 . 2 ILE H H 8.108 0.02 1 12 . 2 ILE HA H 3.894 0.02 1 13 . 2 ILE HB H 1.991 0.02 1 14 . 2 ILE HG13 H 1.307 0.02 1 15 . 2 ILE HG12 H 1.483 0.02 1 16 . 2 ILE HG2 H 0.955 0.02 1 17 . 3 ALA N N 122.701 0.05 1 18 . 3 ALA H H 7.953 0.02 1 19 . 3 ALA HA H 4.057 0.02 1 20 . 3 ALA HB H 1.489 0.02 1 21 . 4 ALA N N 119.926 0.05 1 22 . 4 ALA H H 7.846 0.02 1 23 . 4 ALA HA H 4.229 0.02 1 24 . 4 ALA HB H 1.535 0.02 1 25 . 5 LEU N N 121.318 0.05 1 26 . 5 LEU H H 8.092 0.02 1 27 . 5 LEU HA H 4.148 0.02 1 28 . 5 LEU HB3 H 1.798 0.02 2 29 . 5 LEU HB2 H 1.848 0.02 2 30 . 5 LEU HD2 H 0.923 0.02 2 31 . 6 LYS N N 118.096 0.05 1 32 . 6 LYS H H 8.483 0.02 1 33 . 6 LYS HA H 3.900 0.02 1 34 . 6 LYS HB3 H 1.926 0.02 2 35 . 6 LYS HB2 H 1.970 0.02 2 36 . 6 LYS HG2 H 1.395 0.02 2 37 . 7 GLU N N 120.930 0.05 1 38 . 7 GLU H H 7.918 0.02 1 39 . 7 GLU HA H 4.122 0.02 1 40 . 8 LYS N N 120.696 0.05 1 41 . 8 LYS H H 8.086 0.02 1 42 . 8 LYS HA H 4.155 0.02 1 43 . 8 LYS HB3 H 1.798 0.02 2 44 . 8 LYS HB2 H 1.848 0.02 2 45 . 9 ILE N N 120.203 0.05 1 46 . 9 ILE H H 8.551 0.02 1 47 . 9 ILE HA H 3.689 0.02 1 48 . 9 ILE HB H 1.945 0.02 1 49 . 9 ILE HD1 H 0.845 0.02 1 50 . 9 ILE HG2 H 0.931 0.02 1 51 . 10 ALA N N 121.490 0.05 1 52 . 10 ALA H H 8.157 0.02 1 53 . 10 ALA HA H 4.064 0.02 1 54 . 10 ALA HB H 1.574 0.02 1 55 . 11 ALA N N 118.673 0.05 1 56 . 11 ALA H H 7.922 0.02 1 57 . 11 ALA HA H 4.232 0.02 1 58 . 11 ALA HB H 1.601 0.02 1 59 . 12 LEU N N 121.397 0.05 1 60 . 12 LEU H H 8.357 0.02 1 61 . 12 LEU HA H 4.144 0.02 1 62 . 12 LEU HB3 H 1.918 0.02 2 63 . 12 LEU HG H 1.752 0.02 1 64 . 12 LEU HD1 H 0.917 0.02 2 65 . 12 LEU HD2 H 0.947 0.02 2 66 . 13 LYS N N 118.427 0.05 1 67 . 13 LYS H H 8.705 0.02 1 68 . 13 LYS HA H 3.896 0.02 1 69 . 13 LYS HB3 H 1.989 0.02 2 70 . 13 LYS HB2 H 1.922 0.02 2 71 . 13 LYS HG2 H 1.382 0.02 2 72 . 13 LYS HE2 H 2.887 0.02 2 73 . 14 GLU N N 120.122 0.05 1 74 . 14 GLU H H 7.859 0.02 1 75 . 14 GLU HA H 4.108 0.02 1 76 . 14 GLU HB3 H 2.210 0.02 2 77 . 14 GLU HB2 H 2.104 0.02 2 78 . 14 GLU HG3 H 2.337 0.02 2 79 . 14 GLU HG2 H 2.498 0.02 2 80 . 15 LYS N N 121.003 0.05 1 81 . 15 LYS H H 7.896 0.02 1 82 . 15 LYS HA H 4.156 0.02 1 83 . 15 LYS HB3 H 2.040 0.02 2 84 . 15 LYS HB2 H 2.110 0.02 2 85 . 16 ILE N N 119.615 0.05 1 86 . 16 ILE H H 8.538 0.02 1 87 . 16 ILE HA H 3.661 0.02 1 88 . 16 ILE HB H 1.938 0.02 1 89 . 16 ILE HG2 H 0.844 0.02 1 90 . 17 ALA N N 121.147 0.05 1 91 . 17 ALA H H 8.131 0.02 1 92 . 17 ALA HA H 4.050 0.02 1 93 . 17 ALA HB H 1.538 0.02 1 94 . 18 ALA N N 119.503 0.05 1 95 . 18 ALA H H 7.764 0.02 1 96 . 18 ALA HA H 4.215 0.02 1 97 . 18 ALA HB H 1.580 0.02 1 98 . 19 LEU N N 119.909 0.05 1 99 . 19 LEU H H 8.122 0.02 1 100 . 19 LEU HA H 4.166 0.02 1 101 . 19 LEU HB3 H 2.017 0.02 2 102 . 19 LEU HD1 H 0.916 0.02 2 103 . 20 LYS N N 118.088 0.05 1 104 . 20 LYS H H 8.181 0.02 1 105 . 20 LYS HA H 4.099 0.02 1 106 . 20 LYS HB3 H 1.936 0.02 2 107 . 21 GLU N N 119.118 0.05 1 108 . 21 GLU H H 7.845 0.02 1 109 . 21 GLU HA H 4.211 0.02 1 110 . 21 GLU HB3 H 2.103 0.02 2 111 . 21 GLU HB2 H 2.176 0.02 2 112 . 21 GLU HG3 H 2.362 0.02 2 113 . 21 GLU HG2 H 2.448 0.02 2 stop_ save_