data_26512 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Thermal stability of chicken brain a-spectrin repeat 17: a spectroscopic study ; _BMRB_accession_number 26512 _BMRB_flat_file_name bmr26512.str _Entry_type original _Submission_date 2015-02-17 _Accession_date 2015-02-17 _Entry_origination BMRB _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 Brenner Annette K. . 2 Kieffer Bruno . . 3 Trave Gilles . . 4 Froystein Nils A. . 5 Raae Arnt J. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 3 T1_relaxation 1 T2_relaxation 1 stop_ loop_ _Data_type _Data_type_count "13C chemical shifts" 25 "T1 relaxation values" 65 "T2 relaxation values" 65 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2015-02-25 original author . stop_ _Original_release_date 2015-02-25 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'Thermal stability of chicken brain a-spectrin repeat 17: a spectroscopic study' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 22569754 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Brenner Annette K. . 2 Kieffer Bruno . . 3 Trave Gilles . . 4 Froystein Nils A. . 5 Raae Arnt J. . stop_ _Journal_abbreviation 'J. Biomol. NMR' _Journal_volume 53 _Journal_issue 2 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 71 _Page_last 83 _Year 2012 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name a-spectrin _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label a-spectrin $a-spectrin 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_a-spectrin _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common a-spectrin _Molecular_mass . _Mol_thiol_state 'all free' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 118 _Mol_residue_sequence ; MARGQRLEESLEYQQFVANV EEEEAWINEKMTLVASEDYG DTLAAIQGLLKKHEAFETDF TVHKDRVNDVCANGEDLIKK NNHHVENITAKMKGLKGKVS DLEKAAAQRKAKLDENSA ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 1 MET 2 2 ALA 3 3 ARG 4 4 GLY 5 5 GLN 6 6 ARG 7 7 LEU 8 8 GLU 9 9 GLU 10 10 SER 11 11 LEU 12 12 GLU 13 13 TYR 14 14 GLN 15 15 GLN 16 16 PHE 17 17 VAL 18 18 ALA 19 19 ASN 20 20 VAL 21 21 GLU 22 22 GLU 23 23 GLU 24 24 GLU 25 25 ALA 26 26 TRP 27 27 ILE 28 28 ASN 29 29 GLU 30 30 LYS 31 31 MET 32 32 THR 33 33 LEU 34 34 VAL 35 35 ALA 36 36 SER 37 37 GLU 38 38 ASP 39 39 TYR 40 40 GLY 41 41 ASP 42 42 THR 43 43 LEU 44 44 ALA 45 45 ALA 46 46 ILE 47 47 GLN 48 48 GLY 49 49 LEU 50 50 LEU 51 51 LYS 52 52 LYS 53 53 HIS 54 54 GLU 55 55 ALA 56 56 PHE 57 57 GLU 58 58 THR 59 59 ASP 60 60 PHE 61 61 THR 62 62 VAL 63 63 HIS 64 64 LYS 65 65 ASP 66 66 ARG 67 67 VAL 68 68 ASN 69 69 ASP 70 70 VAL 71 71 CYS 72 72 ALA 73 73 ASN 74 74 GLY 75 75 GLU 76 76 ASP 77 77 LEU 78 78 ILE 79 79 LYS 80 80 LYS 81 81 ASN 82 82 ASN 83 83 HIS 84 84 HIS 85 85 VAL 86 86 GLU 87 87 ASN 88 88 ILE 89 89 THR 90 90 ALA 91 91 LYS 92 92 MET 93 93 LYS 94 94 GLY 95 95 LEU 96 96 LYS 97 97 GLY 98 98 LYS 99 99 VAL 100 100 SER 101 101 ASP 102 102 LEU 103 103 GLU 104 104 LYS 105 105 ALA 106 106 ALA 107 107 ALA 108 108 GLN 109 109 ARG 110 110 LYS 111 111 ALA 112 112 LYS 113 113 LEU 114 114 ASP 115 115 GLU 116 116 ASN 117 117 SER 118 118 ALA stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-03-04 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 18260 R17 100.00 118 100.00 100.00 8.60e-79 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $a-spectrin Chicken 9031 Eukaryota Metazoa Gallus gallus 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 _Details $a-spectrin 'recombinant technology' . Escherichia coli . BL21(DE3) 'pETM12 vector' 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 $a-spectrin 200 uM '[U-100% 13C; U-100% 15N]' D2O 10 % 'natural abundance' H2O 90 % 'natural abundance' 'Sodium phosphate buffer' 20 mM 'natural abundance' NaCl 0.1 M 'natural abundance' DTT 1 mM 'natural abundance' NaN3 0.05 % '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_ 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 'resonance assignment' stop_ _Details . save_ save_CCPNMR _Saveframe_category software _Name CCPNMR _Version . loop_ _Vendor _Address _Electronic_address CCPN . . stop_ loop_ _Task 'data analysis' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model 'Biospin AV600' _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_HNCO_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCO' _Sample_label $sample_1 save_ save_3D_HN(CO)CA_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HN(CO)CA' _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 temperature 298 . K pH 6.8 . pH pressure 1 . atm stop_ save_ save_sample_conditions_2 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units temperature 317 . K pH 6.8 . pH pressure 1 . atm 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 C 13 'methyl protons' ppm 0.00 na indirect . . . 0.251449530 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_ _Software_label $TOPSPIN stop_ loop_ _Experiment_label '3D HNCO' '3D HN(CO)CA' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_2 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name a-spectrin _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 25 25 ALA C C 176.4 . 1 2 25 25 ALA CA C 57.8 . 1 3 33 33 LEU C C 180.1 . 1 4 33 33 LEU CA C 57.8 . 1 5 57 57 GLU C C 180.2 . 1 6 57 57 GLU CA C 60.2 . 1 7 60 60 PHE CA C 56.8 . 1 8 70 70 VAL C C 179.8 . 1 9 70 70 VAL CA C 66 . 1 10 94 94 GLY C C 176.8 . 1 11 94 94 GLY CA C 46.8 . 1 12 99 99 VAL C C 178.2 . 1 13 99 99 VAL CA C 67.6 . 1 stop_ save_ save_assigned_chem_shift_list_2 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Software_label $TOPSPIN stop_ loop_ _Experiment_label '3D HNCO' '3D HN(CO)CA' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_2 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name a-spectrin _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 60 60 PHE C C 176.8 . 1 2 94 94 GLY CA C 53.2 . 1 3 99 99 VAL C C 176.8 . 1 stop_ save_ save_assigned_chem_shift_list_3 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Software_label $TOPSPIN stop_ loop_ _Experiment_label '3D HNCO' '3D HN(CO)CA' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_2 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name a-spectrin _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 25 25 ALA C C 180.8 . 1 2 25 25 ALA CA C 55.1 . 1 3 33 33 LEU C C 177.6 . 1 4 33 33 LEU CA C 52.2 . 1 5 57 57 GLU C C 176.8 . 1 6 57 57 GLU CA C 57 . 1 7 70 70 VAL CA C 58 . 1 8 94 94 GLY CA C 40 . 1 9 99 99 VAL CA C 56.8 . 1 stop_ save_ save_heteronuclear_T1_list_1 _Saveframe_category T1_relaxation _Details . loop_ _Software_label $NMRPipe stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Spectrometer_frequency_1H 600 _T1_coherence_type Sz _T1_value_units s-1 _Mol_system_component_name a-spectrin _Text_data_format . _Text_data . loop_ _T1_ID _Residue_seq_code _Residue_label _Atom_name _T1_value _T1_value_error 1 11 LEU N 1.31 . 2 17 VAL N 1.17 . 3 20 VAL N 0.98 . 4 21 GLU N 1.07 . 5 24 GLU N 0.97 . 6 28 ASN N 0.99 . 7 29 GLU N 1.11 . 8 30 LYS N 0.99 . 9 32 THR N 1.09 . 10 33 LEU N 1.02 . 11 34 VAL N 0.96 . 12 35 ALA N 1.21 . 13 37 GLU N 1.57 . 14 38 ASP N 1.20 . 15 39 TYR N 1.41 . 16 40 GLY N 1.44 . 17 43 LEU N 1.20 . 18 44 ALA N 1.32 . 19 45 ALA N 1.25 . 20 48 GLY N 1.21 . 21 50 LEU N 0.99 . 22 51 LYS N 1.10 . 23 52 LYS N 1.06 . 24 53 HIS N 1.01 . 25 54 GLU N 0.93 . 26 55 ALA N 1.15 . 27 56 PHE N 1.03 . 28 57 GLU N 0.99 . 29 58 THR N 1.10 . 30 59 ASP N 1.06 . 31 60 PHE N 0.97 . 32 61 THR N 1.02 . 33 62 VAL N 1.09 . 34 64 LYS N 1.04 . 35 65 ASP N 1.09 . 36 66 ARG N 1.04 . 37 67 VAL N 0.95 . 38 68 ASN N 1.08 . 39 69 ASP N 1.04 . 40 71 CYS N 1.07 . 41 72 ALA N 1.03 . 42 74 GLY N 0.97 . 43 78 ILE N 1.05 . 44 80 LYS N 1.05 . 45 85 VAL N 1.12 . 46 86 GLU N 1.19 . 47 89 THR N 1.05 . 48 90 ALA N 1.04 . 49 93 LYS N 1.22 . 50 95 LEU N 0.92 . 51 96 LYS N 0.96 . 52 97 GLY N 0.99 . 53 98 LYS N 0.94 . 54 99 VAL N 0.98 . 55 100 SER N 0.92 . 56 103 GLU N 0.93 . 57 104 LYS N 0.96 . 58 105 ALA N 0.91 . 59 106 ALA N 0.97 . 60 107 ALA N 0.95 . 61 108 GLN N 0.94 . 62 110 LYS N 0.97 . 63 111 ALA N 1.04 . 64 112 LYS N 0.95 . 65 114 ASP N 1.06 . stop_ save_ save_heteronuclear_T2_list_1 _Saveframe_category T2_relaxation _Details . loop_ _Software_label $NMRPipe stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Spectrometer_frequency_1H 600 _T2_coherence_type S(+,-) _T2_value_units s-1 _Mol_system_component_name a-spectrin _Text_data_format . _Text_data . loop_ _T2_ID _Residue_seq_code _Residue_label _Atom_name _T2_value _T2_value_error _Rex_value _Rex_error 1 11 LEU N 18.45 . . . 2 17 VAL N 17.89 . . . 3 20 VAL N 16.98 . . . 4 21 GLU N 15.02 . . . 5 24 GLU N 19.65 . . . 6 28 ASN N 18.52 . . . 7 29 GLU N 15.75 . . . 8 30 LYS N 18.22 . . . 9 32 THR N 14.05 . . . 10 33 LEU N 16.89 . . . 11 34 VAL N 18.52 . . . 12 35 ALA N 15.22 . . . 13 37 GLU N 13.55 . . . 14 38 ASP N 16.18 . . . 15 39 TYR N 13.04 . . . 16 40 GLY N 12.63 . . . 17 43 LEU N 18.38 . . . 18 44 ALA N 15.82 . . . 19 45 ALA N 14.97 . . . 20 48 GLY N 17.45 . . . 21 50 LEU N 18.08 . . . 22 51 LYS N 15.82 . . . 23 52 LYS N 17.27 . . . 24 53 HIS N 19.27 . . . 25 54 GLU N 17.18 . . . 26 55 ALA N 16.16 . . . 27 56 PHE N 18.18 . . . 28 57 GLU N 19.80 . . . 29 58 THR N 16.50 . . . 30 59 ASP N 16.78 . . . 31 60 PHE N 19.42 . . . 32 61 THR N 18.12 . . . 33 62 VAL N 16.86 . . . 34 64 LYS N 17.70 . . . 35 65 ASP N 16.37 . . . 36 66 ARG N 18.80 . . . 37 67 VAL N 19.27 . . . 38 68 ASN N 17.33 . . . 39 69 ASP N 16.50 . . . 40 71 CYS N 20.49 . . . 41 72 ALA N 17.36 . . . 42 74 GLY N 19.96 . . . 43 78 ILE N 18.28 . . . 44 80 LYS N 16.67 . . . 45 85 VAL N 19.65 . . . 46 86 GLU N 20.28 . . . 47 89 THR N 19.23 . . . 48 90 ALA N 19.42 . . . 49 93 LYS N 19.01 . . . 50 95 LEU N 18.62 . . . 51 96 LYS N 18.35 . . . 52 97 GLY N 18.94 . . . 53 98 LYS N 18.05 . . . 54 99 VAL N 18.02 . . . 55 100 SER N 18.32 . . . 56 103 GLU N 18.94 . . . 57 104 LYS N 17.64 . . . 58 105 ALA N 18.15 . . . 59 106 ALA N 18.55 . . . 60 107 ALA N 17.73 . . . 61 108 GLN N 18.45 . . . 62 110 LYS N 19.38 . . . 63 111 ALA N 18.59 . . . 64 112 LYS N 18.12 . . . 65 114 ASP N 16.50 . . . stop_ save_