data_25788 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Two-fold symmetric structure of the 18-60 construct of S31N M2 from Influenza A in lipid bilayers ; _BMRB_accession_number 25788 _BMRB_flat_file_name bmr25788.str _Entry_type original _Submission_date 2015-09-01 _Accession_date 2015-09-14 _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 Andreas Loren B. . 2 Reese Marcel . . 3 Eddy Matthew T. . 4 Gelev Vladimir . . 5 Ni 'Qing Zhe' . . 6 Miller Eric A. . 7 Emsley Lyndon . . 8 Pintacuda Guido . . 9 Chou James J. . 10 Griffin Robert G. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 2 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 80 "13C chemical shifts" 250 "15N chemical shifts" 59 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2016-02-24 original BMRB . stop_ _Original_release_date 2016-02-24 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Structure and Mechanism of the Influenza A M218-60 Dimer of Dimers ; _Citation_status published _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 Andreas Loren B. . 2 Reese Marcel . . 3 Eddy Matthew T. . 4 Gelev Vladimir . . 5 Ni 'Qing Zhe' . . 6 Miller Eric A. . 7 Emsley Lyndon . . 8 Pintacuda Guido . . 9 Chou James J. . 10 Griffin Robert G. . stop_ _Journal_abbreviation 'J. Am. Chem. Soc.' _Journal_volume 137 _Journal_issue 47 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 14877 _Page_last 14886 _Year 2015 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'Influenza-A M2' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label entity_1 $matrix_protein_2 entity_2 $matrix_protein_2 entity_3 $matrix_protein_2 entity_4 $matrix_protein_2 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_matrix_protein_2 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common matrix_protein_2 _Molecular_mass . _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 43 _Mol_residue_sequence ; RSNDSSDPLVVAANIIGILH LILWILDRLFFKSIYRFFEH GLK ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 . ARG 2 . SER 3 . ASN 4 . ASP 5 . SER 6 . SER 7 . ASP 8 125 PRO 9 126 LEU 10 127 VAL 11 128 VAL 12 129 ALA 13 130 ALA 14 131 ASN 15 132 ILE 16 133 ILE 17 134 GLY 18 135 ILE 19 136 LEU 20 137 HIS 21 138 LEU 22 139 ILE 23 140 LEU 24 141 TRP 25 142 ILE 26 143 LEU 27 144 ASP 28 145 ARG 29 146 LEU 30 147 PHE 31 148 PHE 32 149 LYS 33 150 SER 34 151 ILE 35 152 TYR 36 . ARG 37 . PHE 38 . PHE 39 . GLU 40 . HIS 41 . GLY 42 . LEU 43 . LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . 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 2KIH 'matrix protein 2' . . . . . stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Strain $matrix_protein_2 'Influenza A virus' 385599 Viruses . 'Influenzavirus A' 'Influenza A virus' A/Udorn/307/1972(H3N2)) 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 $matrix_protein_2 'recombinant technology' . Escherichia coli . PET stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_U-13C-15N_S31N_M2 _Saveframe_category sample _Sample_type membrane _Details ; Per liter, 3 g 13C glucose and 1 g 15N ammonium chloride (CIL), with another 0.5 to 1 g 13C glucose added at induction (150 M IPTG, 18 C) at an OD600 of 0.7 to 0.9. The final OD600 reached 3.5 to 4, after 18 hours of expression at 18 C, 15mg of pure protein. ; loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling 'sodium phosphate' 40 mM 'natural abundance' 'glutamic acid' 30 mM 'natural abundance' 'sodium azide' 3 mM 'natural abundance' 1,2-diphytanoyl-sn-glycero-3-phosphocholine 50 '% w/w' 'natural abundance' $matrix_protein_2 50 '% w/w' [U-13C,15N] H2O 100 % 'natural abundance' stop_ save_ save_U-13C-15N-12C-14N-ILFY-M2 _Saveframe_category sample _Sample_type membrane _Details ; Per liter, 3 g 13C glucose and 1 g 15N ammonium chloride (CIL), with another 0.5 to 1 g 13C glucose added at induction. Cells were grown in 700 mL of media, with 300 mL of media used to dissolve the amino acids. When the OD600 reached 0.7, the reserved 300 mL of media was added, along with amino acids at natural isotopic abundance:23,24 150 mg isoleucine, 150 mg leucine, 50 mg phenylalanine, and 100 mg tyrosine (Sigma-Aldrich). Expression of protein was induced with 150 M IPTG at 18 C as before. The final OD600 reached 3.5 to 4.6, after 18 hours of expression at 18 C, and yielded up to 15 mg of pure protein. ; loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling 'sodium phosphate' 40 mM 'natural abundance' 'glutamic acid' 30 mM 'natural abundance' 'sodium azide' 3 mM 'natural abundance' 1,2-diphytanoyl-sn-glycero-3-phosphocholine 50 '% w/w' 'natural abundance' $matrix_protein_2 50 '% w/w' U-13C,15N-[12C,14N-ILFY] H2O 100 % 'natural abundance' stop_ save_ save_1-13C-Glucose-M2 _Saveframe_category sample _Sample_type membrane _Details ; Preparation was identical to U-13C-15N M2 except that specifically isotopically labeled glucose was used. The labeling pattern resulting from [1,6-13C]-glucose is the same as from [1-13C]-glucose25 (Omicron Biochemicals) except that the [1,6-13C]-glucose results in double the labeling level per site. ; loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling 'sodium phosphate' 40 mM 'natural abundance' 'glutamic acid' 30 mM 'natural abundance' 'sodium azide' 3 mM 'natural abundance' 1,2-diphytanoyl-sn-glycero-3-phosphocholine 50 '% w/w' 'natural abundance' $matrix_protein_2 50 '% w/w' [1-13C]-Glucose H2O 100 % 'natural abundance' stop_ save_ save_1-6-13C2-Glucose-M2 _Saveframe_category sample _Sample_type membrane _Details ; Preparation was identical to U-13C-15N M2 except that specifically isotopically labeled glucose was used. The labeling pattern resulting from [1,6-13C]-glucose is the same as from [1-13C]-glucose25 (Omicron Biochemicals) except that the [1,6-13C]-glucose results in double the labeling level per site. ; loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling 'sodium phosphate' 40 mM 'natural abundance' 'glutamic acid' 30 mM 'natural abundance' 'sodium azide' 3 mM 'natural abundance' 1,2-diphytanoyl-sn-glycero-3-phosphocholine 50 '% w/w' 'natural abundance' $matrix_protein_2 50 '% w/w' [1,6-13C2]-Glucose H2O 100 % 'natural abundance' stop_ save_ save_U-13C-15N-2H-12C-13C2H21H_1-Ile_12C-13Ca-13C-13C2H21H_2-Leu_12C-13C2H21H_2-Val_M2 _Saveframe_category sample _Sample_type membrane _Details ; An overnight culture in 50 mL of 90 % 2H M9, 10 % SOC media was grown from a fresh transformation in BL21-DE3 until an OD600 of 2 to 3. Cells were then pelleted by centrifugation and transferred to 1L of 2H M9 media containing 3 g 2H-13C glucose, 1 g 15N ammonium chloride, salts, and Centrum in 1 L of 99.8 % 2H D2O. The doubling time was 2 hours. At an OD600 of 0.65 to 0.75, ILV precursors were added: 75 mg alpha-keto-butyric acid,26 sodium salt 4-13C 99 %, 3,3,4,4-2H4, 98 % (CIL), and 350 mg of 2-(13C,2H2)methyl-4-(2H3)-acetolactate prepared as described previously. Cells were harvested after 24 to 36 hours, and yielded up to 5 mg of pure protein. This labeling pattern re-sults in methyl groups with isolated 13C1H spin pairs in an otherwise 2H, 12C background at nearby sites. The protein was purified and refolded in 1H buffers, resulting in complete exchange to 1H at exchangeable sites such as the backbone amides. ; loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling 'sodium phosphate' 40 mM 'natural abundance' 'glutamic acid' 30 mM 'natural abundance' 'sodium azide' 3 mM 'natural abundance' 1,2-diphytanoyl-sn-glycero-3-phosphocholine 50 '% w/w' 'aliphatic chain [U-2H]' $matrix_protein_2 50 '% w/w' "U-13C,15N,2H-[12C,13C2H21H 1-Ile, 12C,13Ca,13C',13C2H21H 2-Leu, 12C,13C2H21H 2-Val]" H2O 100 % 'natural abundance' stop_ save_ save_U-13C-15N-2H-13CH3_1-Ile_M2 _Saveframe_category sample _Sample_type membrane _Details 'Precursor was 75 mg of 13C4, 98 %, 3,3-2H2, 98 % alpha-ketobutryic acid, sodium salt.' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling 'sodium phosphate' 40 mM 'natural abundance' 'glutamic acid' 30 mM 'natural abundance' 'sodium azide' 3 mM 'natural abundance' 1,2-diphytanoyl-sn-glycero-3-phosphocholine 50 '% w/w' 'aliphatic chain [U-2H]' $matrix_protein_2 50 '% w/w' 'U-13C,15N,2H-[13CH3 1-Ile]' H2O 100 % 'natural abundance' stop_ save_ save_U-13C-15N-2H-13C2H21H_2-Leu_13C2H21H_2-Val-M2 _Saveframe_category sample _Sample_type membrane _Details 'Precursor was 350 mg of U-13C,2-(2H2)methyl-4-(2H3)-acetolactate' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling 'sodium phosphate' 40 mM 'natural abundance' 'glutamic acid' 30 mM 'natural abundance' 'sodium azide' 3 mM 'natural abundance' 1,2-diphytanoyl-sn-glycero-3-phosphocholine 50 '% w/w' 'aliphatic chain [U-2H]' $matrix_protein_2 50 '% w/w' 'U-13C,15N,2H-[13C2H21H 2-Leu, 13C2H21H 2-Val]' H2O 100 % 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_TALOS+ _Saveframe_category software _Name TALOS+ _Version . loop_ _Vendor _Address _Electronic_address 'Cornilescu, Delaglio and Bax' . . stop_ loop_ _Task 'structure solution' stop_ _Details . save_ save_X-PLOR_NIH _Saveframe_category software _Name X-PLOR_NIH _Version . loop_ _Vendor _Address _Electronic_address 'Schwieters, Kuszewski, Tjandra and Clore' . . stop_ loop_ _Task 'structure solution' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 800 _Details . save_ save_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 900 _Details . save_ save_spectrometer_3 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 1000 _Details . save_ save_spectrometer_4 _Saveframe_category NMR_spectrometer _Manufacturer FBML _Model 'Cambridge Instruments' _Field_strength 750 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_ZF-TEDOR_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D ZF-TEDOR' _Sample_label $U-13C-15N_S31N_M2 save_ save_3D_ZF-TEDOR-RFDR_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D ZF-TEDOR-RFDR' _Sample_label $U-13C-15N_S31N_M2 save_ save_2D_HN_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D HN' _Sample_label $U-13C-15N_S31N_M2 save_ save_3D_(H)_CaNH_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D (H) CaNH' _Sample_label $U-13C-15N-12C-14N-ILFY-M2 save_ save_2D_PAR_5 _Saveframe_category NMR_applied_experiment _Experiment_name '2D PAR' _Sample_label $U-13C-15N_S31N_M2 save_ save_2D_PDSD_6 _Saveframe_category NMR_applied_experiment _Experiment_name '2D PDSD' _Sample_label $U-13C-15N_S31N_M2 save_ save_2D_ZF-TEDOR_7 _Saveframe_category NMR_applied_experiment _Experiment_name '2D ZF-TEDOR' _Sample_label $1-6-13C2-Glucose-M2 save_ save_2D_RFDR_8 _Saveframe_category NMR_applied_experiment _Experiment_name '2D RFDR' _Sample_label $1-6-13C2-Glucose-M2 save_ save_3D_1H-1H_RFDR_9 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-1H RFDR' _Sample_label $U-13C-15N-12C-14N-ILFY-M2 save_ save_4D_HCHHCH_10 _Saveframe_category NMR_applied_experiment _Experiment_name '4D HCHHCH' _Sample_label $U-13C-15N-12C-14N-ILFY-M2 save_ save_2D_C-H_11 _Saveframe_category NMR_applied_experiment _Experiment_name '2D C-H' _Sample_label $U-13C-15N-2H-12C-13C2H21H_1-Ile_12C-13Ca-13C-13C2H21H_2-Leu_12C-13C2H21H_2-Val_M2 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 7.8 . pH temperature 298 5 K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference_1 _Saveframe_category chemical_shift_reference _Details 'DSS scale using adamantane as a secondary reference.' 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_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D ZF-TEDOR' '3D ZF-TEDOR-RFDR' '2D HN' '3D (H) CaNH' '2D RFDR' '2D C-H' stop_ loop_ _Sample_label $U-13C-15N_S31N_M2 $U-13C-15N-12C-14N-ILFY-M2 $1-6-13C2-Glucose-M2 $U-13C-15N-2H-12C-13C2H21H_1-Ile_12C-13Ca-13C-13C2H21H_2-Leu_12C-13C2H21H_2-Val_M2 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name entity_1 _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 1 125 8 PRO C C 177.886 . . 2 125 8 PRO CA C 65.367 . . 3 125 8 PRO CB C 32.689 . . 4 125 8 PRO CG C 27.468 . . 5 125 8 PRO CD C 50.728 . . 6 125 8 PRO N N 135.180 . . 7 126 9 LEU H H 8.691 . . 8 126 9 LEU C C 178.187 . . 9 126 9 LEU CA C 58.123 . . 10 126 9 LEU CB C 40.799 . . 11 126 9 LEU CG C 28.138 . . 12 126 9 LEU CD1 C 25.860 . . 13 126 9 LEU CD2 C 23.301 . . 14 126 9 LEU N N 115.733 . . 15 127 10 VAL H H 7.510 . . 16 127 10 VAL HG2 H 1.161 . . 17 127 10 VAL C C 178.610 . . 18 127 10 VAL CA C 66.092 . . 19 127 10 VAL CB C 31.866 . . 20 127 10 VAL CG1 C 21.637 . . 21 127 10 VAL CG2 C 23.136 . . 22 127 10 VAL N N 119.338 . . 23 128 11 VAL H H 8.189 . . 24 128 11 VAL HG2 H 1.120 . . 25 128 11 VAL C C 178.273 . . 26 128 11 VAL CA C 66.650 . . 27 128 11 VAL CB C 31.325 . . 28 128 11 VAL CG1 C 21.898 . . 29 128 11 VAL CG2 C 22.483 . . 30 128 11 VAL N N 120.038 . . 31 129 12 ALA H H 8.439 . . 32 129 12 ALA C C 179.401 . . 33 129 12 ALA CA C 55.148 . . 34 129 12 ALA CB C 18.492 . . 35 129 12 ALA N N 118.367 . . 36 130 13 ALA H H 8.428 . . 37 130 13 ALA C C 176.912 . . 38 130 13 ALA CA C 55.001 . . 39 130 13 ALA CB C 18.605 . . 40 130 13 ALA N N 118.963 . . 41 131 14 ASN H H 7.686 . . 42 131 14 ASN C C 175.078 . . 43 131 14 ASN CA C 54.740 . . 44 131 14 ASN CB C 40.594 . . 45 131 14 ASN CG C 177.469 . . 46 131 14 ASN N N 110.836 . . 47 131 14 ASN ND2 N 112.920 . . 48 132 15 ILE H H 7.712 . . 49 132 15 ILE HG12 H 0.854 . . 50 132 15 ILE HG13 H 0.854 . . 51 132 15 ILE C C 177.746 . . 52 132 15 ILE CA C 61.350 . . 53 132 15 ILE CB C 41.265 . . 54 132 15 ILE CG1 C 26.260 . . 55 132 15 ILE CG2 C 17.200 . . 56 132 15 ILE CD1 C 14.284 . . 57 132 15 ILE N N 110.420 . . 58 133 16 ILE H H 7.621 . . 59 133 16 ILE HG12 H 1.006 . . 60 133 16 ILE HG13 H 1.006 . . 61 133 16 ILE C C 175.337 . . 62 133 16 ILE CA C 64.273 . . 63 133 16 ILE CB C 37.633 . . 64 133 16 ILE CG1 C 28.877 . . 65 133 16 ILE CG2 C 17.187 . . 66 133 16 ILE CD1 C 14.621 . . 67 133 16 ILE N N 121.299 . . 68 134 17 GLY H H 9.154 . . 69 134 17 GLY C C 175.979 . . 70 134 17 GLY CA C 47.617 . . 71 134 17 GLY N N 109.835 . . 72 135 18 ILE H H 8.265 . . 73 135 18 ILE HG12 H 1.108 . . 74 135 18 ILE HG13 H 1.108 . . 75 135 18 ILE C C 176.873 . . 76 135 18 ILE CA C 64.134 . . 77 135 18 ILE CB C 37.126 . . 78 135 18 ILE CG1 C 28.797 . . 79 135 18 ILE CG2 C 17.725 . . 80 135 18 ILE CD1 C 12.693 . . 81 135 18 ILE N N 121.438 . . 82 136 19 LEU H H 8.464 . . 83 136 19 LEU C C 177.412 . . 84 136 19 LEU CA C 57.605 . . 85 136 19 LEU CB C 41.688 . . 86 136 19 LEU CG C 26.801 . . 87 136 19 LEU CD1 C 26.074 . . 88 136 19 LEU CD2 C 24.674 . . 89 136 19 LEU N N 118.641 . . 90 137 20 HIS H H 8.599 . . 91 137 20 HIS HE2 H 14.889 . . 92 137 20 HIS C C 175.564 . . 93 137 20 HIS CA C 59.266 . . 94 137 20 HIS CB C 30.327 . . 95 137 20 HIS CG C 136.279 . . 96 137 20 HIS CD2 C 119.147 . . 97 137 20 HIS CE1 C 136.880 . . 98 137 20 HIS N N 116.550 . . 99 137 20 HIS ND1 N 251.069 . . 100 137 20 HIS NE2 N 173.470 . . 101 138 21 LEU H H 7.981 . . 102 138 21 LEU HG H 1.057 . . 103 138 21 LEU C C 178.731 . . 104 138 21 LEU CA C 58.418 . . 105 138 21 LEU CB C 42.313 . . 106 138 21 LEU CG C 26.415 . . 107 138 21 LEU CD1 C 26.498 . . 108 138 21 LEU CD2 C 22.509 . . 109 138 21 LEU N N 116.334 . . 110 139 22 ILE H H 8.289 . . 111 139 22 ILE HG12 H 1.047 . . 112 139 22 ILE HG13 H 1.047 . . 113 139 22 ILE C C 175.907 . . 114 139 22 ILE CA C 60.425 . . 115 139 22 ILE CB C 37.178 . . 116 139 22 ILE CG1 C 30.230 . . 117 139 22 ILE CG2 C 19.651 . . 118 139 22 ILE CD1 C 13.536 . . 119 139 22 ILE N N 114.269 . . 120 140 23 LEU H H 8.518 . . 121 140 23 LEU HG H 1.091 . . 122 140 23 LEU C C 178.534 . . 123 140 23 LEU CA C 58.505 . . 124 140 23 LEU CB C 41.306 . . 125 140 23 LEU CG C 26.929 . . 126 140 23 LEU CD1 C 25.008 . . 127 140 23 LEU CD2 C 24.440 . . 128 140 23 LEU N N 126.307 . . 129 141 24 TRP H H 9.799 . . 130 141 24 TRP HE1 H 10.826 . . 131 141 24 TRP C C 178.577 . . 132 141 24 TRP CA C 60.713 . . 133 141 24 TRP CB C 28.216 . . 134 141 24 TRP CG C 111.799 . . 135 141 24 TRP CD1 C 125.059 . . 136 141 24 TRP CD2 C 129.327 . . 137 141 24 TRP CE2 C 138.728 . . 138 141 24 TRP CE3 C 119.699 . . 139 141 24 TRP CZ2 C 114.040 . . 140 141 24 TRP CZ3 C 121.655 . . 141 141 24 TRP CH2 C 124.358 . . 142 141 24 TRP N N 121.039 . . 143 141 24 TRP NE1 N 128.847 . . 144 142 25 ILE H H 8.380 . . 145 142 25 ILE HG12 H 1.015 . . 146 142 25 ILE HG13 H 1.015 . . 147 142 25 ILE C C 178.361 . . 148 142 25 ILE CA C 64.227 . . 149 142 25 ILE CB C 37.941 . . 150 142 25 ILE CG1 C 28.792 . . 151 142 25 ILE CG2 C 18.555 . . 152 142 25 ILE CD1 C 14.180 . . 153 142 25 ILE N N 116.764 . . 154 143 26 LEU H H 8.858 . . 155 143 26 LEU C C 179.930 . . 156 143 26 LEU CA C 57.709 . . 157 143 26 LEU CB C 41.673 . . 158 143 26 LEU CG C 26.471 . . 159 143 26 LEU CD1 C 23.212 . . 160 143 26 LEU CD2 C 22.593 . . 161 143 26 LEU N N 119.319 . . 162 144 27 ASP H H 9.446 . . 163 144 27 ASP C C 178.506 . . 164 144 27 ASP CA C 57.865 . . 165 144 27 ASP CB C 42.759 . . 166 144 27 ASP CG C 180.189 . . 167 144 27 ASP N N 120.965 . . 168 145 28 ARG H H 8.569 . . 169 145 28 ARG C C 178.483 . . 170 145 28 ARG CA C 57.778 . . 171 145 28 ARG CB C 29.567 . . 172 145 28 ARG CG C 25.899 . . 173 145 28 ARG CD C 42.762 . . 174 145 28 ARG CZ C 159.631 . . 175 145 28 ARG N N 116.733 . . 176 145 28 ARG NE N 84.466 . . 177 146 29 LEU H H 8.133 . . 178 146 29 LEU C C 177.521 . . 179 146 29 LEU CA C 56.325 . . 180 146 29 LEU CB C 43.000 . . 181 146 29 LEU CG C 26.670 . . 182 146 29 LEU CD1 C 25.481 . . 183 146 29 LEU CD2 C 22.935 . . 184 146 29 LEU N N 114.549 . . 185 147 30 PHE H H 7.946 . . 186 147 30 PHE C C 175.633 . . 187 147 30 PHE CA C 58.408 . . 188 147 30 PHE CB C 41.525 . . 189 147 30 PHE CD1 C 131.845 . . 190 147 30 PHE CD2 C 131.845 . . 191 147 30 PHE N N 112.643 . . 192 148 31 PHE H H 7.763 . . 193 148 31 PHE C C 175.834 . . 194 148 31 PHE CA C 60.950 . . 195 148 31 PHE CB C 39.071 . . 196 148 31 PHE CD1 C 131.807 . . 197 148 31 PHE CD2 C 131.807 . . 198 148 31 PHE N N 116.558 . . 199 149 32 LYS H H 10.295 . . 200 149 32 LYS C C 175.590 . . 201 149 32 LYS CA C 56.723 . . 202 149 32 LYS CB C 30.395 . . 203 149 32 LYS CG C 25.198 . . 204 149 32 LYS CD C 29.343 . . 205 149 32 LYS CE C 42.456 . . 206 149 32 LYS N N 120.261 . . 207 149 32 LYS NZ N 32.214 . . 208 150 33 SER H H 9.688 . . 209 150 33 SER C C 174.425 . . 210 150 33 SER CA C 60.620 . . 211 150 33 SER CB C 66.910 . . 212 150 33 SER N N 110.950 . . 213 151 34 ILE H H 10.356 . . 214 151 34 ILE HG12 H 0.922 . . 215 151 34 ILE HG13 H 0.922 . . 216 151 34 ILE C C 175.964 . . 217 151 34 ILE CA C 65.844 . . 218 151 34 ILE CB C 38.232 . . 219 151 34 ILE CG1 C 28.515 . . 220 151 34 ILE CG2 C 17.221 . . 221 151 34 ILE CD1 C 13.782 . . 222 151 34 ILE N N 126.206 . . 223 152 35 TYR N N 118.049 . . stop_ save_ save_assigned_chem_shift_list_1_2 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D ZF-TEDOR' '3D ZF-TEDOR-RFDR' '2D HN' '3D (H) CaNH' '2D RFDR' '2D C-H' stop_ loop_ _Sample_label $U-13C-15N_S31N_M2 $U-13C-15N-12C-14N-ILFY-M2 $1-6-13C2-Glucose-M2 $U-13C-15N-2H-12C-13C2H21H_1-Ile_12C-13Ca-13C-13C2H21H_2-Leu_12C-13C2H21H_2-Val_M2 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name entity_2 _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 1 224 7 ASP H H 8.474 . . 2 224 7 ASP C C 175.000 . . 3 224 7 ASP CA C 51.985 . . 4 224 7 ASP CB C 41.865 . . 5 224 7 ASP CG C 179.075 . . 6 224 7 ASP N N 124.149 . . 7 225 8 PRO C C 177.512 . . 8 225 8 PRO CA C 65.323 . . 9 225 8 PRO CB C 32.625 . . 10 225 8 PRO CG C 27.416 . . 11 225 8 PRO CD C 50.667 . . 12 225 8 PRO N N 136.194 . . 13 226 9 LEU H H 8.638 . . 14 226 9 LEU C C 178.309 . . 15 226 9 LEU CA C 57.858 . . 16 226 9 LEU CB C 41.287 . . 17 226 9 LEU CG C 26.501 . . 18 226 9 LEU CD1 C 25.285 . . 19 226 9 LEU CD2 C 22.575 . . 20 226 9 LEU N N 116.185 . . 21 227 10 VAL H H 7.429 . . 22 227 10 VAL HG2 H 1.114 . . 23 227 10 VAL C C 177.680 . . 24 227 10 VAL CA C 66.618 . . 25 227 10 VAL CB C 31.523 . . 26 227 10 VAL CG1 C 22.444 . . 27 227 10 VAL CG2 C 21.605 . . 28 227 10 VAL N N 119.065 . . 29 228 11 VAL H H 7.984 . . 30 228 11 VAL HG2 H 1.239 . . 31 228 11 VAL C C 178.699 . . 32 228 11 VAL CA C 65.627 . . 33 228 11 VAL CB C 31.658 . . 34 228 11 VAL CG1 C 21.540 . . 35 228 11 VAL CG2 C 22.663 . . 36 228 11 VAL N N 118.746 . . 37 229 12 ALA H H 8.525 . . 38 229 12 ALA C C 178.638 . . 39 229 12 ALA CA C 55.372 . . 40 229 12 ALA CB C 18.438 . . 41 229 12 ALA N N 119.881 . . 42 230 13 ALA H H 8.364 . . 43 230 13 ALA C C 178.264 . . 44 230 13 ALA CA C 55.377 . . 45 230 13 ALA CB C 18.704 . . 46 230 13 ALA N N 117.892 . . 47 231 14 ASN H H 8.725 . . 48 231 14 ASN C C 177.489 . . 49 231 14 ASN CA C 58.010 . . 50 231 14 ASN CB C 39.892 . . 51 231 14 ASN CG C 174.810 . . 52 231 14 ASN N N 115.603 . . 53 231 14 ASN ND2 N 115.328 . . 54 232 15 ILE H H 8.688 . . 55 232 15 ILE HG12 H 0.954 . . 56 232 15 ILE HG13 H 0.954 . . 57 232 15 ILE C C 177.215 . . 58 232 15 ILE CA C 65.574 . . 59 232 15 ILE CB C 37.178 . . 60 232 15 ILE CG1 C 29.611 . . 61 232 15 ILE CG2 C 17.644 . . 62 232 15 ILE CD1 C 14.116 . . 63 232 15 ILE N N 118.775 . . 64 233 16 ILE H H 8.472 . . 65 233 16 ILE HG12 H 0.856 . . 66 233 16 ILE HG13 H 0.856 . . 67 233 16 ILE C C 177.650 . . 68 233 16 ILE CA C 66.000 . . 69 233 16 ILE CB C 37.145 . . 70 233 16 ILE CG1 C 30.240 . . 71 233 16 ILE CG2 C 19.624 . . 72 233 16 ILE CD1 C 13.610 . . 73 233 16 ILE N N 120.164 . . 74 234 17 GLY H H 8.718 . . 75 234 17 GLY C C 175.208 . . 76 234 17 GLY CA C 47.952 . . 77 234 17 GLY N N 106.809 . . 78 235 18 ILE H H 8.621 . . 79 235 18 ILE HG12 H 0.991 . . 80 235 18 ILE HG13 H 0.991 . . 81 235 18 ILE C C 177.200 . . 82 235 18 ILE CA C 66.085 . . 83 235 18 ILE CB C 37.595 . . 84 235 18 ILE CG1 C 30.693 . . 85 235 18 ILE CG2 C 17.960 . . 86 235 18 ILE CD1 C 14.213 . . 87 235 18 ILE N N 121.901 . . 88 236 19 LEU H H 8.797 . . 89 236 19 LEU HG H 1.085 . . 90 236 19 LEU C C 177.284 . . 91 236 19 LEU CA C 58.132 . . 92 236 19 LEU CB C 41.264 . . 93 236 19 LEU CG C 26.467 . . 94 236 19 LEU CD1 C 24.726 . . 95 236 19 LEU CD2 C 24.437 . . 96 236 19 LEU N N 119.929 . . 97 237 20 HIS H H 8.721 . . 98 237 20 HIS HE2 H 12.151 . . 99 237 20 HIS C C 177.331 . . 100 237 20 HIS CA C 62.320 . . 101 237 20 HIS CB C 31.270 . . 102 237 20 HIS CG C 135.819 . . 103 237 20 HIS CD2 C 117.215 . . 104 237 20 HIS CE1 C 135.748 . . 105 237 20 HIS N N 118.864 . . 106 237 20 HIS ND1 N 251.566 . . 107 237 20 HIS NE2 N 166.796 . . 108 238 21 LEU H H 8.275 . . 109 238 21 LEU HG H 0.855 . . 110 238 21 LEU C C 178.271 . . 111 238 21 LEU CA C 58.255 . . 112 238 21 LEU CB C 40.425 . . 113 238 21 LEU CG C 26.509 . . 114 238 21 LEU CD1 C 21.965 . . 115 238 21 LEU CD2 C 26.042 . . 116 238 21 LEU N N 120.531 . . 117 239 22 ILE H H 8.983 . . 118 239 22 ILE HG12 H 0.930 . . 119 239 22 ILE HG13 H 0.930 . . 120 239 22 ILE C C 177.405 . . 121 239 22 ILE CA C 65.543 . . 122 239 22 ILE CB C 37.249 . . 123 239 22 ILE CG1 C 29.603 . . 124 239 22 ILE CG2 C 17.207 . . 125 239 22 ILE CD1 C 13.869 . . 126 239 22 ILE N N 118.698 . . 127 240 23 LEU H H 8.702 . . 128 240 23 LEU C C 179.334 . . 129 240 23 LEU CA C 58.130 . . 130 240 23 LEU CB C 40.791 . . 131 240 23 LEU CG C 26.996 . . 132 240 23 LEU CD1 C 26.207 . . 133 240 23 LEU CD2 C 23.212 . . 134 240 23 LEU N N 118.843 . . 135 241 24 TRP H H 8.402 . . 136 241 24 TRP HE1 H 11.128 . . 137 241 24 TRP C C 177.814 . . 138 241 24 TRP CA C 62.610 . . 139 241 24 TRP CB C 27.421 . . 140 241 24 TRP CG C 111.077 . . 141 241 24 TRP CD1 C 127.471 . . 142 241 24 TRP CD2 C 130.876 . . 143 241 24 TRP CE2 C 138.835 . . 144 241 24 TRP CE3 C 120.962 . . 145 241 24 TRP CZ2 C 114.895 . . 146 241 24 TRP CZ3 C 120.905 . . 147 241 24 TRP CH2 C 123.768 . . 148 241 24 TRP N N 122.683 . . 149 241 24 TRP NE1 N 130.688 . . 150 242 25 ILE H H 9.044 . . 151 242 25 ILE HG12 H 1.000 . . 152 242 25 ILE HG13 H 1.000 . . 153 242 25 ILE HG2 H 0.941 . . 154 242 25 ILE C C 178.059 . . 155 242 25 ILE CA C 65.807 . . 156 242 25 ILE CB C 37.802 . . 157 242 25 ILE CG1 C 29.431 . . 158 242 25 ILE CG2 C 17.115 . . 159 242 25 ILE CD1 C 14.391 . . 160 242 25 ILE N N 119.630 . . 161 243 26 LEU H H 8.912 . . 162 243 26 LEU C C 177.586 . . 163 243 26 LEU CA C 58.062 . . 164 243 26 LEU CB C 41.336 . . 165 243 26 LEU N N 117.887 . . 166 244 27 ASP N N 117.765 . . stop_ save_