data_19254 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; NMR assignments and backbone amide relaxation data (T1-T2) for the complex of the Y48A mutant of the FimH adhesin carbohydrate-binding domain with heptyl-mannose ; _BMRB_accession_number 19254 _BMRB_flat_file_name bmr19254.str _Entry_type original _Submission_date 2013-05-22 _Accession_date 2013-05-22 _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 Vanwetswinkel Sophie . . 2 'Van Nuland' N. A. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 T1_relaxation 1 T2_relaxation 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 188 "15N chemical shifts" 170 "T1 relaxation values" 133 "T2 relaxation values" 130 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2014-03-04 update author 'update entry citation' 2014-02-13 original author 'original release' stop_ loop_ _Related_BMRB_accession_number _Relationship 19066 'FimH adhesin carbohydrate-binding domain' 19255 'FimH Y48A mutant' 19256 'FimH -heptyl-mannose complex' stop_ _Original_release_date 2013-05-22 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Study of the Structural and Dynamic Effects in the FimH Adhesin upon alfa-D-Heptyl Mannose Binding ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 24476493 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Vanwetswinkel Sophie . . 2 Volkov Alexander N. . 3 Sterckx Yann G.J. . 4 Garcia-Pino Abel . . 5 Buts Lieven . . 6 Vranken Wim F. . 7 Bouckaert Julie . . 8 Roy Rene . . 9 Wyns Lode . . 10 'van Nuland' Nico A.J. . stop_ _Journal_abbreviation 'J. Med. Chem.' _Journal_name_full 'Journal of medicinal chemistry' _Journal_volume 57 _Journal_issue 4 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 1416 _Page_last 1427 _Year 2014 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'FimH Y48A mutant - Heptyl-mannose complex' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'FIMH Y48A mutant' $FIMH_Y48A_mutant heptyl-mannose $entity_KGM stop_ _System_molecular_weight 16904.8881 _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state 'all disulfide bound' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_FIMH_Y48A_mutant _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common FIMH_Y48A_mutant _Molecular_mass 16904.8881 _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 158 _Mol_residue_sequence ; FACKTANGTAIPIGGGSANV YVNLAPVVNVGQNLVVDLST QIFCHNDAPETITDYVTLQR GSAYGGVLSNFSGTVKYSGS SYPFPTTSETPRVVYNSRTD KPWPVALYLTPVSSAGGVAI KAGSLIAVLILRQTNNYNSD DFQFVWNIYANNDVVVPT ; loop_ _Residue_seq_code _Residue_label 1 PHE 2 ALA 3 CYS 4 LYS 5 THR 6 ALA 7 ASN 8 GLY 9 THR 10 ALA 11 ILE 12 PRO 13 ILE 14 GLY 15 GLY 16 GLY 17 SER 18 ALA 19 ASN 20 VAL 21 TYR 22 VAL 23 ASN 24 LEU 25 ALA 26 PRO 27 VAL 28 VAL 29 ASN 30 VAL 31 GLY 32 GLN 33 ASN 34 LEU 35 VAL 36 VAL 37 ASP 38 LEU 39 SER 40 THR 41 GLN 42 ILE 43 PHE 44 CYS 45 HIS 46 ASN 47 ASP 48 ALA 49 PRO 50 GLU 51 THR 52 ILE 53 THR 54 ASP 55 TYR 56 VAL 57 THR 58 LEU 59 GLN 60 ARG 61 GLY 62 SER 63 ALA 64 TYR 65 GLY 66 GLY 67 VAL 68 LEU 69 SER 70 ASN 71 PHE 72 SER 73 GLY 74 THR 75 VAL 76 LYS 77 TYR 78 SER 79 GLY 80 SER 81 SER 82 TYR 83 PRO 84 PHE 85 PRO 86 THR 87 THR 88 SER 89 GLU 90 THR 91 PRO 92 ARG 93 VAL 94 VAL 95 TYR 96 ASN 97 SER 98 ARG 99 THR 100 ASP 101 LYS 102 PRO 103 TRP 104 PRO 105 VAL 106 ALA 107 LEU 108 TYR 109 LEU 110 THR 111 PRO 112 VAL 113 SER 114 SER 115 ALA 116 GLY 117 GLY 118 VAL 119 ALA 120 ILE 121 LYS 122 ALA 123 GLY 124 SER 125 LEU 126 ILE 127 ALA 128 VAL 129 LEU 130 ILE 131 LEU 132 ARG 133 GLN 134 THR 135 ASN 136 ASN 137 TYR 138 ASN 139 SER 140 ASP 141 ASP 142 PHE 143 GLN 144 PHE 145 VAL 146 TRP 147 ASN 148 ILE 149 TYR 150 ALA 151 ASN 152 ASN 153 ASP 154 VAL 155 VAL 156 VAL 157 PRO 158 THR 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 UNP A2IC68_ECOLX A2IC68 . . . . . stop_ save_ ############# # Ligands # ############# save_KGM _Saveframe_category ligand _Mol_type "non-polymer (NON-POLYMER)" _Name_common 'Heptyl Alpha-D-mannopyrannoside' _BMRB_code KGM _PDB_code KGM _Molecular_mass 278.342 _Mol_charge 0 _Mol_paramagnetic . _Mol_aromatic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons O6 O6 O . 0 . ? C6 C6 C . 0 . ? C5 C5 C . 0 . ? C10 C10 C . 0 . ? O5 O5 O . 0 . ? C4 C4 C . 0 . ? O4 O4 O . 0 . ? C3 C3 C . 0 . ? O3 O3 O . 0 . ? C2 C2 C . 0 . ? O2 O2 O . 0 . ? C1 C1 C . 0 . ? O1 O1 O . 0 . ? C7 C7 C . 0 . ? C8 C8 C . 0 . ? C9 C9 C . 0 . ? C13 C13 C . 0 . ? C12 C12 C . 0 . ? C11 C11 C . 0 . ? H131 H131 H . 0 . ? H132 H132 H . 0 . ? H133 H133 H . 0 . ? H121 H121 H . 0 . ? H122 H122 H . 0 . ? H111 H111 H . 0 . ? H112 H112 H . 0 . ? H101 H101 H . 0 . ? H102 H102 H . 0 . ? H91C H91C H . 0 . ? H92C H92C H . 0 . ? H81C H81C H . 0 . ? H82C H82C H . 0 . ? H71C H71C H . 0 . ? H72C H72C H . 0 . ? H1 H1 H . 0 . ? H2 H2 H . 0 . ? HA HA H . 0 . ? H3 H3 H . 0 . ? HB HB H . 0 . ? H4 H4 H . 0 . ? HC HC H . 0 . ? H5 H5 H . 0 . ? H61C H61C H . 0 . ? H62C H62C H . 0 . ? H6 H6 H . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name SING C13 C12 ? ? SING C12 C11 ? ? SING C11 C10 ? ? SING C10 C9 ? ? SING C9 C8 ? ? SING C8 C7 ? ? SING C7 O1 ? ? SING O1 C1 ? ? SING C1 C2 ? ? SING C1 O5 ? ? SING C2 O2 ? ? SING C2 C3 ? ? SING C3 O3 ? ? SING C3 C4 ? ? SING C4 O4 ? ? SING C4 C5 ? ? SING C5 O5 ? ? SING C5 C6 ? ? SING C6 O6 ? ? SING C13 H131 ? ? SING C13 H132 ? ? SING C13 H133 ? ? SING C12 H121 ? ? SING C12 H122 ? ? SING C11 H111 ? ? SING C11 H112 ? ? SING C10 H101 ? ? SING C10 H102 ? ? SING C9 H91C ? ? SING C9 H92C ? ? SING C8 H81C ? ? SING C8 H82C ? ? SING C7 H71C ? ? SING C7 H72C ? ? SING C1 H1 ? ? SING C2 H2 ? ? SING O2 HA ? ? SING C3 H3 ? ? SING O3 HB ? ? SING C4 H4 ? ? SING O4 HC ? ? SING C5 H5 ? ? SING C6 H61C ? ? SING C6 H62C ? ? SING O6 H6 ? ? stop_ _Mol_thiol_state . _Sequence_homology_query_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Strain _Details $FIMH_Y48A_mutant Enterobacteria 562 Bacteria . Escherichia coli J96 'N-TERMINAL LECTIN DOMAIN, RESIDUES 22-179' 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 $FIMH_Y48A_mutant 'recombinant technology' 'Escherichia coli' . . BL21(DE3) pET24a $entity_KGM 'chemical synthesis' . . . . . stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details 'FimH_Y48A 0.0011 M, sodium phosphate 0.020000 M, sodium chloride 0.100000 M' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $FIMH_Y48A_mutant 0.0011 M '[U-100% 15N]' 'sodium phosphate' 0.02 M 'natural abundance' 'sodium chloride' 0.1 M 'natural abundance' $entity_KGM 1.3 mM 'natural abundance' H2O 93 % 'natural abundance' D2O 7 % 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_CcpNmr_Analysis _Saveframe_category software _Name ANALYSIS _Version 2.1 _Details . save_ save_nmrPipe_any _Saveframe_category software _Name NMRPipe _Version any _Details . save_ save_NMRDraw _Saveframe_category software _Name NMRDraw _Version . loop_ _Vendor _Address _Electronic_address 'Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax' . . stop_ loop_ _Task 'spectrum analysis' 'spectrum display' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model Direct-Drive _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_1H-15N_NOESY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-15N NOESY' _Sample_label $sample_1 save_ save_2D_1H-15N_HSQC_(inverse_recovery)_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC (inverse recovery)' _Sample_label $sample_1 save_ save_2D_1H-15N_HSQC_(CPMG)_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC (CPMG)' _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details 'pH [6.0], temp [298], pressure [1.0], ionStrength [0.0]' loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 0.12 . M pH 6.000 . pH pressure 1.000 . atm temperature 298.000 . 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 HDO H 1 proton ppm 4.7 internal indirect . . . 1 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 _Saveframe_category assigned_chemical_shifts _Details 'Origin nmrStar file /ebi/msd/pdb_root/Processing/prepare/3zpd/ebi/EBI-55996.csh' loop_ _Experiment_label '2D 1H-15N HSQC' '3D 1H-15N NOESY' 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 'FIMH Y48A mutant' _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 2 ALA H H 6.72675 0 . 2 2 2 ALA N N 129.55102 0 . 3 3 3 CYS H H 7.79390 0 . 4 3 3 CYS N N 112.24906 0 . 5 4 4 LYS H H 10.01731 0 . 6 4 4 LYS N N 121.47945 0 . 7 5 5 THR H H 8.80318 0 . 8 5 5 THR N N 109.53710 0 . 9 6 6 ALA H H 8.66805 0 . 10 6 6 ALA N N 123.49563 0 . 11 7 7 ASN H H 8.12819 0 . 12 7 7 ASN HD21 H 7.09795 0 . 13 7 7 ASN HD22 H 7.83071 0 . 14 7 7 ASN N N 112.91246 0 . 15 7 7 ASN ND2 N 112.49515 0 . 16 8 8 GLY H H 8.08212 0 . 17 8 8 GLY N N 107.78406 0 . 18 9 9 THR H H 7.76840 0 . 19 9 9 THR N N 118.29281 0 . 20 10 10 ALA H H 8.71829 0 . 21 10 10 ALA N N 127.84147 0 . 22 11 11 ILE H H 8.47735 0 . 23 11 11 ILE N N 120.39858 0 . 24 13 13 ILE H H 7.76242 0 . 25 13 13 ILE N N 116.30523 0 . 26 14 14 GLY H H 8.27253 0 . 27 14 14 GLY N N 116.28567 0 . 28 15 15 GLY H H 7.62580 0 . 29 15 15 GLY N N 108.85820 0 . 30 16 16 GLY H H 7.59343 0 . 31 16 16 GLY N N 104.22186 0 . 32 17 17 SER H H 8.33949 0 . 33 17 17 SER N N 115.59443 0 . 34 18 18 ALA H H 8.54808 0 . 35 18 18 ALA N N 122.31717 0 . 36 19 19 ASN H H 8.67153 0 . 37 19 19 ASN HD21 H 6.95132 0 . 38 19 19 ASN HD22 H 7.39617 0 . 39 19 19 ASN N N 119.09728 0 . 40 19 19 ASN ND2 N 112.41569 0 . 41 20 20 VAL H H 9.49909 0 . 42 20 20 VAL N N 124.17321 0 . 43 21 21 TYR H H 8.84878 0 . 44 21 21 TYR N N 128.43481 0 . 45 22 22 VAL H H 8.73715 0 . 46 22 22 VAL N N 116.12323 0 . 47 23 23 ASN H H 8.89896 0 . 48 23 23 ASN HD21 H 7.09430 0 . 49 23 23 ASN HD22 H 7.65106 0 . 50 23 23 ASN N N 124.25760 0 . 51 23 23 ASN ND2 N 114.46751 0 . 52 24 24 LEU H H 9.09496 0 . 53 24 24 LEU N N 124.58135 0 . 54 25 25 ALA H H 7.90067 0 . 55 25 25 ALA N N 128.92936 0 . 56 27 27 VAL H H 7.34738 0 . 57 27 27 VAL N N 118.12124 0 . 58 28 28 VAL H H 8.62849 0 . 59 28 28 VAL N N 126.56877 0 . 60 29 29 ASN H H 8.10306 0 . 61 29 29 ASN HD21 H 6.82567 0 . 62 29 29 ASN HD22 H 7.32658 0 . 63 29 29 ASN N N 121.60292 0 . 64 29 29 ASN ND2 N 111.05696 0 . 65 30 30 VAL H H 8.61862 0 . 66 30 30 VAL N N 119.47779 0 . 67 31 31 GLY H H 8.79128 0 . 68 31 31 GLY N N 115.24651 0 . 69 32 32 GLN H H 8.11655 0 . 70 32 32 GLN HE21 H 6.92257 0 . 71 32 32 GLN HE22 H 7.65743 0 . 72 32 32 GLN N N 119.99495 0 . 73 32 32 GLN NE2 N 112.70102 0 . 74 33 33 ASN H H 8.51340 0 . 75 33 33 ASN HD21 H 6.89062 0 . 76 33 33 ASN HD22 H 7.46992 0 . 77 33 33 ASN N N 116.76852 0 . 78 33 33 ASN ND2 N 111.69581 0 . 79 34 34 LEU H H 9.83780 0 . 80 34 34 LEU N N 126.42075 0 . 81 35 35 VAL H H 8.62799 0 . 82 35 35 VAL N N 127.52760 0 . 83 36 36 VAL H H 8.94085 0 . 84 36 36 VAL N N 129.17984 0 . 85 37 37 ASP H H 8.98073 0 . 86 37 37 ASP N N 126.53692 0 . 87 38 38 LEU H H 8.50509 0 . 88 38 38 LEU N N 125.36540 0 . 89 39 39 SER H H 8.71952 0 . 90 39 39 SER N N 120.31867 0 . 91 40 40 THR H H 6.66159 0 . 92 40 40 THR N N 105.60599 0 . 93 41 41 GLN H H 7.58530 0 . 94 41 41 GLN HE21 H 6.80809 0 . 95 41 41 GLN HE22 H 7.47859 0 . 96 41 41 GLN N N 117.31673 0 . 97 41 41 GLN NE2 N 112.32923 0 . 98 42 42 ILE H H 7.19861 0 . 99 42 42 ILE N N 118.06001 0 . 100 43 43 PHE H H 8.55566 0 . 101 43 43 PHE N N 123.44902 0 . 102 44 44 CYS H H 9.21756 0 . 103 44 44 CYS N N 118.10247 0 . 104 45 45 HIS H H 7.42411 0 . 105 45 45 HIS N N 112.69739 0 . 106 46 46 ASN H H 7.78523 0 . 107 46 46 ASN N N 118.73702 0 . 108 47 47 ASP H H 9.42453 0 . 109 47 47 ASP N N 123.88724 0 . 110 48 48 ALA H H 9.88928 0 . 111 48 48 ALA N N 124.30316 0 . 112 50 50 GLU H H 9.00215 0 . 113 50 50 GLU N N 118.95185 0 . 114 51 51 THR H H 7.60711 0 . 115 51 51 THR N N 106.47418 0 . 116 52 52 ILE H H 8.41499 0.00824 . 117 52 52 ILE N N 122.97538 0.00728 . 118 53 53 THR H H 8.46453 0 . 119 53 53 THR N N 123.80841 0 . 120 54 54 ASP H H 6.84686 0 . 121 54 54 ASP N N 124.37107 0 . 122 55 55 TYR H H 8.64182 0.00338 . 123 55 55 TYR N N 121.25942 0.03680 . 124 56 56 VAL H H 9.25935 0 . 125 56 56 VAL N N 121.47371 0 . 126 57 57 THR H H 9.64564 0 . 127 57 57 THR N N 123.65212 0 . 128 58 58 LEU H H 7.42380 0 . 129 58 58 LEU N N 120.38492 0 . 130 59 59 GLN H H 9.21656 0 . 131 59 59 GLN HE21 H 6.59354 0 . 132 59 59 GLN HE22 H 7.22308 0 . 133 59 59 GLN N N 130.94001 0 . 134 59 59 GLN NE2 N 108.62127 0 . 135 60 60 ARG H H 7.12996 0 . 136 60 60 ARG HE H 7.13714 0 . 137 60 60 ARG N N 112.70820 0 . 138 60 60 ARG NE N 84.33444 0 . 139 61 61 GLY H H 9.31419 0 . 140 61 61 GLY N N 112.70062 0 . 141 62 62 SER H H 9.15504 0 . 142 62 62 SER N N 122.31910 0 . 143 63 63 ALA H H 8.12545 0 . 144 63 63 ALA N N 125.90309 0 . 145 64 64 TYR H H 7.64045 0 . 146 64 64 TYR N N 116.13553 0 . 147 65 65 GLY H H 8.93962 0 . 148 65 65 GLY N N 108.39936 0 . 149 66 66 GLY H H 9.64479 0 . 150 66 66 GLY N N 114.64713 0 . 151 67 67 VAL H H 7.66892 0 . 152 67 67 VAL N N 119.71390 0 . 153 68 68 LEU H H 6.57313 0 . 154 68 68 LEU N N 118.77115 0 . 155 69 69 SER H H 7.88211 0 . 156 69 69 SER N N 109.30434 0 . 157 70 70 ASN H H 8.10547 0 . 158 70 70 ASN HD21 H 7.33928 0 . 159 70 70 ASN HD22 H 7.89919 0 . 160 70 70 ASN N N 114.27093 0 . 161 70 70 ASN ND2 N 116.97080 0 . 162 71 71 PHE H H 7.86793 0 . 163 71 71 PHE N N 116.99563 0 . 164 72 72 SER H H 9.19566 0 . 165 72 72 SER N N 115.36688 0 . 166 73 73 GLY H H 8.93463 0 . 167 73 73 GLY N N 109.68365 0 . 168 74 74 THR H H 8.69480 0 . 169 74 74 THR N N 111.86074 0 . 170 75 75 VAL H H 9.57498 0 . 171 75 75 VAL N N 119.75318 0 . 172 76 76 LYS H H 9.48374 0 . 173 76 76 LYS N N 128.25656 0 . 174 77 77 TYR H H 9.36052 0 . 175 77 77 TYR N N 125.55995 0 . 176 78 78 SER H H 8.87141 0 . 177 78 78 SER N N 125.31728 0 . 178 79 79 GLY H H 8.72177 0 . 179 79 79 GLY N N 105.22046 0 . 180 80 80 SER H H 7.50326 0 . 181 80 80 SER N N 117.19923 0 . 182 81 81 SER H H 8.30643 0 . 183 81 81 SER N N 118.02933 0 . 184 82 82 TYR H H 9.21535 0 . 185 82 82 TYR N N 123.76452 0 . 186 84 84 PHE H H 7.50369 0 . 187 84 84 PHE N N 119.39808 0 . 188 86 86 THR H H 8.34254 0 . 189 86 86 THR N N 113.67125 0 . 190 87 87 THR H H 8.77847 0 . 191 87 87 THR N N 110.27300 0 . 192 88 88 SER H H 7.68867 0 . 193 88 88 SER N N 116.25114 0 . 194 89 89 GLU H H 8.40548 0 . 195 89 89 GLU N N 124.55261 0 . 196 90 90 THR H H 7.56053 0 . 197 90 90 THR N N 116.99307 0 . 198 92 92 ARG H H 8.20401 0 . 199 92 92 ARG HE H 6.88276 0 . 200 92 92 ARG N N 117.52330 0 . 201 92 92 ARG NE N 83.60225 0 . 202 93 93 VAL H H 9.23568 0 . 203 93 93 VAL N N 121.21695 0 . 204 94 94 VAL H H 8.47735 0 . 205 94 94 VAL N N 125.33790 0 . 206 95 95 TYR H H 9.14305 0 . 207 95 95 TYR N N 126.41740 0 . 208 96 96 ASN H H 9.15367 0 . 209 96 96 ASN HD21 H 6.86182 0 . 210 96 96 ASN HD22 H 7.62100 0 . 211 96 96 ASN N N 124.30448 0 . 212 96 96 ASN ND2 N 110.45875 0 . 213 97 97 SER H H 7.40635 0 . 214 97 97 SER N N 109.65279 0 . 215 98 98 ARG H H 9.42436 0 . 216 98 98 ARG N N 128.88217 0 . 217 99 99 THR H H 8.31773 0 . 218 99 99 THR N N 116.79467 0 . 219 100 100 ASP H H 8.46808 0 . 220 100 100 ASP N N 126.17244 0 . 221 101 101 LYS H H 9.39072 0 . 222 101 101 LYS N N 124.70455 0 . 223 103 103 TRP H H 8.32867 0 . 224 103 103 TRP HE1 H 9.04644 0 . 225 103 103 TRP N N 128.39914 0 . 226 103 103 TRP NE1 N 128.07575 0 . 227 105 105 VAL H H 8.09143 0 . 228 105 105 VAL N N 125.75851 0 . 229 106 106 ALA H H 8.36651 0 . 230 106 106 ALA N N 125.91757 0 . 231 107 107 LEU H H 8.64936 0 . 232 107 107 LEU N N 120.14294 0 . 233 108 108 TYR H H 8.95075 0 . 234 108 108 TYR N N 123.33578 0 . 235 109 109 LEU H H 9.01397 0 . 236 109 109 LEU N N 124.96660 0 . 237 110 110 THR H H 9.64406 0 . 238 110 110 THR N N 120.40145 0 . 239 112 112 VAL H H 8.09132 0 . 240 112 112 VAL N N 118.33551 0 . 241 113 113 SER H H 8.31478 0 . 242 113 113 SER N N 114.86049 0 . 243 114 114 SER H H 7.73010 0 . 244 114 114 SER N N 112.80350 0 . 245 115 115 ALA H H 7.73922 0 . 246 115 115 ALA N N 126.06817 0 . 247 116 116 GLY H H 8.11857 0 . 248 116 116 GLY N N 106.52736 0 . 249 117 117 GLY H H 8.64779 0 . 250 117 117 GLY N N 111.76920 0 . 251 118 118 VAL H H 8.62331 0 . 252 118 118 VAL N N 122.61808 0 . 253 119 119 ALA H H 9.10129 0 . 254 119 119 ALA N N 134.67730 0 . 255 120 120 ILE H H 7.62826 0 . 256 120 120 ILE N N 115.64396 0 . 257 121 121 LYS H H 8.41122 0 . 258 121 121 LYS N N 125.64257 0 . 259 122 122 ALA H H 9.83224 0 . 260 122 122 ALA N N 129.02456 0 . 261 123 123 GLY H H 9.05948 0 . 262 123 123 GLY N N 111.86906 0 . 263 124 124 SER H H 8.07658 0 . 264 124 124 SER N N 115.03589 0 . 265 125 125 LEU H H 8.24843 0 . 266 125 125 LEU N N 123.97913 0 . 267 126 126 ILE H H 9.07937 0 . 268 126 126 ILE N N 119.64020 0 . 269 127 127 ALA H H 7.71383 0 . 270 127 127 ALA N N 120.28178 0 . 271 128 128 VAL H H 8.54795 0 . 272 128 128 VAL N N 119.18673 0 . 273 129 129 LEU H H 9.40483 0 . 274 129 129 LEU N N 128.61097 0 . 275 130 130 ILE H H 9.07419 0 . 276 130 130 ILE N N 121.84137 0 . 277 131 131 LEU H H 9.36227 0 . 278 131 131 LEU N N 134.72223 0 . 279 132 132 ARG H H 9.05250 0 . 280 132 132 ARG HE H 8.57562 0 . 281 132 132 ARG N N 130.08930 0 . 282 132 132 ARG NE N 86.43386 0 . 283 133 133 GLN H H 9.46054 0 . 284 133 133 GLN HE21 H 6.07695 0 . 285 133 133 GLN HE22 H 6.96268 0 . 286 133 133 GLN N N 132.87027 0 . 287 133 133 GLN NE2 N 110.23914 0 . 288 134 134 THR H H 8.88185 0 . 289 134 134 THR N N 117.90172 0 . 290 135 135 ASN H H 9.76333 0 . 291 135 135 ASN HD21 H 7.65051 0 . 292 135 135 ASN HD22 H 7.77677 0 . 293 135 135 ASN N N 117.68470 0 . 294 135 135 ASN ND2 N 116.80679 0 . 295 136 136 ASN H H 8.32100 0 . 296 136 136 ASN HD21 H 7.16831 0 . 297 136 136 ASN HD22 H 7.76391 0 . 298 136 136 ASN N N 115.96942 0 . 299 136 136 ASN ND2 N 112.54330 0 . 300 137 137 TYR H H 9.10843 0.00239 . 301 137 137 TYR N N 123.39985 0.01215 . 302 138 138 ASN H H 9.44239 0 . 303 138 138 ASN HD21 H 6.97154 0 . 304 138 138 ASN HD22 H 7.71952 0 . 305 138 138 ASN N N 121.65048 0 . 306 138 138 ASN ND2 N 111.25276 0 . 307 139 139 SER H H 7.85187 0 . 308 139 139 SER N N 111.79638 0 . 309 140 140 ASP H H 7.73834 0 . 310 140 140 ASP N N 124.29157 0 . 311 141 141 ASP H H 6.63185 0 . 312 141 141 ASP N N 121.75488 0 . 313 142 142 PHE H H 8.99704 0 . 314 142 142 PHE N N 123.37396 0 . 315 143 143 GLN H H 8.81555 0 . 316 143 143 GLN HE21 H 6.60288 0 . 317 143 143 GLN HE22 H 7.76804 0 . 318 143 143 GLN N N 118.15389 0 . 319 143 143 GLN NE2 N 112.30259 0 . 320 144 144 PHE H H 9.93641 0 . 321 144 144 PHE N N 127.37878 0 . 322 145 145 VAL H H 8.78337 0 . 323 145 145 VAL N N 125.30404 0 . 324 146 146 TRP H H 8.76164 0 . 325 146 146 TRP HE1 H 7.08843 0 . 326 146 146 TRP N N 126.80193 0 . 327 146 146 TRP NE1 N 122.69595 0 . 328 147 147 ASN H H 9.01233 0 . 329 147 147 ASN HD21 H 6.30259 0 . 330 147 147 ASN HD22 H 6.67607 0 . 331 147 147 ASN N N 125.00531 0 . 332 147 147 ASN ND2 N 110.64046 0 . 333 148 148 ILE H H 9.23452 0 . 334 148 148 ILE N N 124.60849 0 . 335 149 149 TYR H H 9.31919 0 . 336 149 149 TYR N N 127.29395 0 . 337 150 150 ALA H H 8.56573 0 . 338 150 150 ALA N N 122.33946 0 . 339 151 151 ASN H H 9.13364 0 . 340 151 151 ASN HD21 H 6.55451 0 . 341 151 151 ASN HD22 H 7.38329 0 . 342 151 151 ASN N N 120.11389 0 . 343 151 151 ASN ND2 N 109.44680 0 . 344 152 152 ASN H H 7.39191 0 . 345 152 152 ASN HD21 H 7.17088 0 . 346 152 152 ASN HD22 H 7.24360 0 . 347 152 152 ASN N N 110.95367 0 . 348 152 152 ASN ND2 N 114.96764 0 . 349 153 153 ASP H H 8.47456 0 . 350 153 153 ASP N N 117.20144 0 . 351 154 154 VAL H H 8.83495 0 . 352 154 154 VAL N N 121.20648 0 . 353 155 155 VAL H H 8.70919 0 . 354 155 155 VAL N N 128.33778 0 . 355 156 156 VAL H H 8.49982 0 . 356 156 156 VAL N N 126.99049 0 . 357 158 158 THR H H 7.71492 0 . 358 158 158 THR N N 118.89152 0 . stop_ save_ save_heteronuclear_T1_list_1 _Saveframe_category T1_relaxation _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Spectrometer_frequency_1H 600 _T1_coherence_type Nz _T1_value_units ms _Mol_system_component_name 'FIMH Y48A mutant' _Text_data_format . _Text_data . loop_ _T1_ID _Residue_seq_code _Residue_label _Atom_name _T1_value _T1_value_error 1 2 ALA N 815.64464 20.05605 2 3 CYS N 935.91103 20.04468 3 4 LYS N 893.74550 11.72869 4 5 THR N 900.22993 10.12632 5 6 ALA N 976.76349 26.46685 6 7 ASN N 841.13713 20.26928 7 8 GLY N 859.11597 20.87730 8 9 THR N 993.36873 19.30649 9 10 ALA N 919.43480 11.03271 10 11 ILE N 894.89886 22.01249 11 13 ILE N 922.67343 39.96646 12 14 GLY N 821.08582 33.81433 13 15 GLY N 879.51258 21.75873 14 16 GLY N 955.98609 21.53678 15 17 SER N 973.43925 35.60817 16 19 ASN N 1015.59015 24.90159 17 20 VAL N 994.71899 20.42811 18 21 TYR N 1099.31906 20.42085 19 22 VAL N 940.72393 19.70014 20 23 ASN N 1123.09100 22.50534 21 24 LEU N 1019.32370 15.95864 22 25 ALA N 984.72807 8.62651 23 27 VAL N 959.29473 24.31659 24 28 VAL N 888.15974 46.42628 25 29 ASN N 996.62058 21.77153 26 30 VAL N 1126.83476 23.03453 27 31 GLY N 898.12447 21.30881 28 32 GLN N 897.06677 25.73665 29 33 ASN N 950.52829 16.02139 30 34 LEU N 891.30292 28.64136 31 35 VAL N 860.73309 26.33382 32 36 VAL N 912.10642 24.12995 33 37 ASP N 972.54312 28.09075 34 39 SER N 1109.71440 17.75398 35 40 THR N 984.10792 25.46613 36 42 ILE N 1019.83779 22.44197 37 43 PHE N 889.29761 22.60848 38 44 CYS N 912.50639 25.15110 39 45 HIS N 881.96829 24.73576 40 46 ASN N 885.25298 38.77757 41 48 ALA N 1120.96785 36.18337 42 50 GLU N 903.78410 14.57658 43 51 THR N 977.41456 35.99796 44 52 ILE N 904.65645 29.76542 45 53 THR N 936.72833 24.08254 46 54 ASP N 950.37285 40.16172 47 55 TYR N 898.66681 29.34367 48 56 VAL N 980.93764 46.05401 49 57 THR N 900.29843 11.22758 50 58 LEU N 835.60797 33.59762 51 59 GLN N 868.98171 20.78773 52 60 ARG N 959.75357 40.30743 53 61 GLY N 952.00417 32.34296 54 62 SER N 1020.04948 42.33267 55 64 TYR N 891.98013 11.66926 56 65 GLY N 875.13453 36.50482 57 66 GLY N 867.58726 23.79145 58 67 VAL N 922.03403 22.52056 59 68 LEU N 898.65111 18.31618 60 69 SER N 950.92751 20.48500 61 70 ASN N 1019.68368 34.33015 62 71 PHE N 910.52643 15.09703 63 72 SER N 922.14816 20.86853 64 73 GLY N 944.00197 10.54375 65 74 THR N 919.76973 7.60512 66 75 VAL N 886.97119 17.92646 67 76 LYS N 895.74423 17.94608 68 77 TYR N 886.40811 36.11412 69 78 SER N 885.92203 45.32905 70 79 GLY N 848.25799 43.16238 71 80 SER N 1000.34466 15.26369 72 81 SER N 939.97380 17.44699 73 82 TYR N 917.75757 21.04847 74 84 PHE N 1123.12749 20.57051 75 86 THR N 834.18642 27.04340 76 87 THR N 788.83372 25.19376 77 88 SER N 954.06022 17.84502 78 89 GLU N 1036.41658 42.05392 79 92 ARG N 1031.26876 12.27420 80 93 VAL N 898.13208 23.37011 81 95 TYR N 965.49629 42.77202 82 97 SER N 991.44450 44.51548 83 98 ARG N 984.31248 64.68188 84 99 THR N 817.81080 20.13523 85 100 ASP N 850.63781 42.43980 86 101 LYS N 933.11022 19.67740 87 103 TRP N 933.08214 23.36836 88 106 ALA N 896.86127 14.40499 89 107 LEU N 927.20046 18.32865 90 110 THR N 967.01863 16.84624 91 112 VAL N 820.57253 24.26399 92 113 SER N 841.35553 35.97827 93 114 SER N 903.94017 20.74094 94 115 ALA N 810.90746 8.41766 95 116 GLY N 845.24596 8.16867 96 117 GLY N 884.70621 16.69879 97 118 VAL N 1049.27688 16.54787 98 119 ALA N 888.02431 19.11762 99 120 ILE N 1049.16385 20.47186 100 121 LYS N 911.77309 17.21293 101 122 ALA N 1155.82186 24.60816 102 123 GLY N 892.46435 22.77518 103 124 SER N 909.14287 23.73372 104 125 LEU N 955.67828 31.13744 105 126 ILE N 1003.57746 32.80571 106 127 ALA N 864.34618 17.07225 107 128 VAL N 982.21498 31.30250 108 129 LEU N 924.33945 17.64085 109 130 ILE N 1000.54915 57.72464 110 131 LEU N 891.98652 32.54528 111 132 ARG N 862.14451 32.88137 112 133 GLN N 843.84774 15.11202 113 134 THR N 907.52662 25.98490 114 135 ASN N 915.35507 15.17063 115 136 ASN N 895.44655 39.98166 116 137 TYR N 1043.17174 31.31905 117 138 ASN N 1105.79481 50.12667 118 139 SER N 719.22472 25.11674 119 140 ASP N 1028.47615 29.35569 120 141 ASP N 979.22607 26.86751 121 143 GLN N 1057.86831 25.01323 122 144 PHE N 919.75683 33.14998 123 145 VAL N 924.29609 20.43033 124 146 TRP N 989.15464 37.43974 125 148 ILE N 891.75576 26.89566 126 149 TYR N 1074.78477 32.11053 127 151 ASN N 923.99851 17.81051 128 152 ASN N 1011.02313 30.55870 129 153 ASP N 959.61194 15.80690 130 154 VAL N 875.02826 22.56067 131 155 VAL N 925.62504 19.29704 132 156 VAL N 910.87305 32.94907 133 158 THR N 1026.87720 17.94823 stop_ save_ save_heteronuclear_T2_list_1 _Saveframe_category T2_relaxation _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Spectrometer_frequency_1H 600 _T2_coherence_type Nz _T2_value_units ms _Mol_system_component_name 'FIMH Y48A mutant' _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 2 ALA N 45.06896 0.84484 . . 2 3 CYS N 47.25785 1.46324 . . 3 4 LYS N 48.97378 1.35055 . . 4 5 THR N 44.95286 1.11180 . . 5 6 ALA N 45.52070 0.84820 . . 6 7 ASN N 54.92236 1.96904 . . 7 8 GLY N 50.46233 1.66993 . . 8 9 THR N 49.73071 0.71911 . . 9 10 ALA N 57.55827 0.84890 . . 10 11 ILE N 51.04894 1.16124 . . 11 13 ILE N 43.75158 1.65564 . . 12 14 GLY N 47.76310 1.65073 . . 13 15 GLY N 39.11720 0.65751 . . 14 16 GLY N 40.73288 1.34929 . . 15 17 SER N 43.73827 1.03451 . . 16 19 ASN N 44.79398 0.66784 . . 17 20 VAL N 44.60561 1.44678 . . 18 21 TYR N 44.64149 1.75466 . . 19 22 VAL N 44.85263 1.06992 . . 20 23 ASN N 40.62520 1.91367 . . 21 24 LEU N 44.69108 1.44246 . . 22 25 ALA N 44.02790 1.50484 . . 23 27 VAL N 40.68736 1.02841 . . 24 28 VAL N 41.73956 2.02232 . . 25 29 ASN N 38.94089 2.08795 . . 26 30 VAL N 44.15299 0.57065 . . 27 31 GLY N 49.85093 1.52252 . . 28 32 GLN N 45.65101 1.09117 . . 29 33 ASN N 52.23925 1.46795 . . 30 34 LEU N 50.60406 1.60874 . . 31 35 VAL N 43.50028 2.07646 . . 32 36 VAL N 48.18399 0.99323 . . 33 37 ASP N 48.33443 1.49092 . . 34 39 SER N 33.91585 1.21231 . . 35 40 THR N 36.84568 1.46478 . . 36 42 ILE N 48.16893 0.63540 . . 37 43 PHE N 46.05906 2.79959 . . 38 44 CYS N 43.64564 0.82941 . . 39 45 HIS N 42.74251 1.16400 . . 40 46 ASN N 43.74948 1.64594 . . 41 48 ALA N 36.76666 0.89447 . . 42 50 GLU N 44.83707 0.74643 . . 43 51 THR N 45.70653 0.88863 . . 44 52 ILE N 46.44183 1.85394 . . 45 53 THR N 42.57261 1.25138 . . 46 54 ASP N 48.31455 1.66452 . . 47 55 TYR N 45.43804 1.04874 . . 48 56 VAL N 48.28405 1.63197 . . 49 57 THR N 44.58563 1.18042 . . 50 58 LEU N 44.72224 1.56680 . . 51 59 GLN N 46.99149 2.50855 . . 52 61 GLY N 40.75806 1.16901 . . 53 62 SER N 38.44644 1.07441 . . 54 64 TYR N 48.64820 1.04575 . . 55 65 GLY N 44.69152 1.41272 . . 56 66 GLY N 40.25381 0.82615 . . 57 67 VAL N 40.06509 1.49396 . . 58 68 LEU N 45.29026 0.90782 . . 59 69 SER N 40.59687 0.72239 . . 60 70 ASN N 43.46292 1.42025 . . 61 71 PHE N 42.84410 1.46632 . . 62 72 SER N 47.22846 0.88424 . . 63 73 GLY N 49.96632 1.63425 . . 64 74 THR N 47.08565 0.42069 . . 65 75 VAL N 43.86167 1.81232 . . 66 76 LYS N 44.25372 1.33230 . . 67 77 TYR N 44.70104 1.10815 . . 68 78 SER N 44.67311 2.38223 . . 69 79 GLY N 51.33966 1.68008 . . 70 80 SER N 44.96539 1.14367 . . 71 81 SER N 48.90204 1.39137 . . 72 82 TYR N 46.69691 1.29238 . . 73 84 PHE N 41.02944 1.08975 . . 74 86 THR N 40.96679 1.28531 . . 75 87 THR N 50.09817 0.81360 . . 76 88 SER N 53.56944 0.93021 . . 77 89 GLU N 48.74021 1.49636 . . 78 92 ARG N 47.84231 0.96616 . . 79 93 VAL N 48.45004 1.12273 . . 80 95 TYR N 46.61568 1.24958 . . 81 97 SER N 33.68176 1.60806 . . 82 98 ARG N 40.58474 1.67527 . . 83 99 THR N 42.13615 0.53467 . . 84 100 ASP N 50.24421 1.40101 . . 85 101 LYS N 50.21493 1.07271 . . 86 103 TRP N 44.12792 1.64676 . . 87 106 ALA N 46.07719 0.68902 . . 88 107 LEU N 44.39819 1.67375 . . 89 110 THR N 46.01105 1.37432 . . 90 112 VAL N 36.59829 1.67113 . . 91 113 SER N 45.86199 1.11440 . . 92 114 SER N 48.41475 1.17138 . . 93 115 ALA N 53.29149 0.77750 . . 94 116 GLY N 65.81764 0.80388 . . 95 117 GLY N 57.64609 2.00754 . . 96 118 VAL N 51.17141 1.17075 . . 97 119 ALA N 46.03276 1.57962 . . 98 121 LYS N 45.49785 1.93403 . . 99 122 ALA N 38.35753 1.07223 . . 100 123 GLY N 43.09008 1.55385 . . 101 124 SER N 40.86078 1.48746 . . 102 125 LEU N 41.15848 1.54853 . . 103 127 ALA N 40.78289 1.04955 . . 104 128 VAL N 40.40117 1.07994 . . 105 129 LEU N 45.13139 1.14267 . . 106 130 ILE N 40.67261 1.55384 . . 107 131 LEU N 46.09043 1.64380 . . 108 132 ARG N 42.41790 0.81640 . . 109 133 GLN N 41.69258 1.64432 . . 110 134 THR N 42.48303 1.03844 . . 111 135 ASN N 43.98107 1.01824 . . 112 136 ASN N 43.01578 1.35042 . . 113 137 TYR N 39.97518 1.06477 . . 114 138 ASN N 41.60293 1.53076 . . 115 139 SER N 116.05473 10.55395 . . 116 140 ASP N 34.23056 1.07769 . . 117 141 ASP N 47.59511 0.95561 . . 118 143 GLN N 44.93099 2.03124 . . 119 144 PHE N 51.27031 2.06119 . . 120 145 VAL N 40.95234 0.97429 . . 121 146 TRP N 47.38393 2.05281 . . 122 148 ILE N 43.91325 1.57584 . . 123 149 TYR N 37.33700 1.74841 . . 124 151 ASN N 46.53635 1.19795 . . 125 152 ASN N 48.47740 1.13660 . . 126 153 ASP N 44.50782 1.12758 . . 127 154 VAL N 47.86365 1.19102 . . 128 155 VAL N 45.19536 1.67419 . . 129 156 VAL N 47.62257 1.29310 . . 130 158 THR N 65.20600 1.04197 . . stop_ save_