data_6144 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; The pyrophosphate cage: the structure of the nisin/lipid II complex provides a blueprint for novel antibiotics ; _BMRB_accession_number 6144 _BMRB_flat_file_name bmr6144.str _Entry_type original _Submission_date 2004-03-16 _Accession_date 2004-03-17 _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 Hsu Shang-Te D. . 2 'de Kruijff' Ben . . 3 Tischenko Eugene . . 4 Kaptein Robert . . 5 Breukink Eefjan . . 6 Bonvin Alexandre M. . 7 Lutters Mandy A. . 8 'van Nuland' Nico A. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 172 "13C chemical shifts" 67 "15N chemical shifts" 32 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2008-08-06 update BMRB 'updating non-standard residues' 2004-10-13 original author 'original release' stop_ loop_ _Related_BMRB_accession_number _Relationship 6145 '3LII in DMSO' 6146 'nisin/3LII complex' stop_ save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'The nisin-lipid II complex reveals a pyrophosphate cage that provides a blueprint for novel antibiotics' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 15361862 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Hsu Shang-Te D. . 2 Breukink Eefjan . . 3 Tischenko Eugene . . 4 Lutters Mandy A. . 5 'de Kruijff' Ben . . 6 Kaptein Robert . . 7 Bonvin Alexandre M. . 8 'van Nuland' Nico A. . stop_ _Journal_abbreviation 'Nat. Struct. Mol. Biol.' _Journal_volume 11 _Journal_issue 10 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 963 _Page_last 967 _Year 2004 _Details . save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Hsu ST, Breukink E, de Kruijff B, Kaptein R, Bonvin AM, van Nuland NA. Biochemistry. 2002 Jun 18;41(24):7670-6. ; _Citation_title 'Mapping the targeted membrane pore formation mechanism by solution NMR: the nisin Z and lipid II interaction in SDS micelles.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 12056898 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Hsu Shang-Te T. . 2 Breukink Eefjan . . 3 'de Kruijff' Ben . . 4 Kaptein Robert . . 5 Bonvin 'Alexandre M J J' M. . 6 'van Nuland' 'Nico A J' A. . stop_ _Journal_abbreviation Biochemistry _Journal_name_full Biochemistry _Journal_volume 41 _Journal_issue 24 _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first 7670 _Page_last 7676 _Year 2002 _Details ; Nisin is an example of type-A lantibiotics that contain cyclic lanthionine rings and unusual dehydrated amino acids. Among the numerous pore-forming antimicrobial peptides, type-A lantibiotics form an unique family of post-translationally modified peptides. Via the recognition of cell wall precursor lipid II, nisin has the capacity to form pores against Gram-positive bacteria with an extremely high activity in the nanomolar (nM) range. Here we report a high-resolution NMR spectroscopy study of nisin/lipid II interactions in SDS micelles as a model membrane system in order to elucidate the mechanism of molecular recognition at residue level. The binding to lipid II was studied through (15)N-(1)H HSQC titration, backbone amide proton temperature coefficient analysis, and heteronuclear (15)N[(1)H]-NOE relaxation dynamics experiments. Upon the addition of lipid II, significant changes were monitored in the N-terminal part of nisin. An extremely low amide proton temperature coefficient (Delta delta/Delta T) was found for the amide proton of Ala3 (> -0.1 ppb/K) in the complex form. This suggests tight hydrogen bonding and/or isolation from the bulk solvent for this residue. Large chemical shift perturbations were also observed in the first two rings. In contrast, the C-terminal part of nisin was almost unaffected. This part of the molecule remains flexible and solvent-exposed. On the basis of our results, a multistep pore-forming mechanism is proposed. The N-terminal part of nisin first binds to lipid II, and a subsequent structural rearrangement takes place. The C-terminal part of nisin is possibly responsible for the activation of the pore formation. In light of the emerging antibiotic resistance problems, an understanding of the specific recognition mechanism of nisin with lipid II at the residue specific level may therefore aid in the development of novel antibiotics. ; save_ ################################## # Molecular system description # ################################## save_system_nisin _Saveframe_category molecular_system _Mol_system_name nisin _Abbreviation_common nisin _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'nisin monomer' $nisin stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state heterodimer _System_paramagnetic no _System_thiol_state 'all other bound' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_nisin _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'nisin Z' _Abbreviation_common nisin _Molecular_mass . _Mol_thiol_state 'all other bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 34 _Mol_residue_sequence ; IXXIXLCXPGCKXGALMGCN MKXAXCNCSIHVXK ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 1 ILE 2 2 DBU 3 3 DAL 4 4 ILE 5 5 DHA 6 6 LEU 7 7 CYS 8 8 ABA 9 9 PRO 10 10 GLY 11 11 CYS 12 12 LYS 13 13 ABA 14 14 GLY 15 15 ALA 16 16 LEU 17 17 MET 18 18 GLY 19 19 CYS 20 20 ASN 21 21 MET 22 22 LYS 23 23 ABA 24 24 ALA 25 25 ABA 26 26 CYS 27 27 ASN 28 28 CYS 29 29 SER 30 30 ILE 31 31 HIS 32 32 VAL 33 33 DHA 34 34 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ ###################### # Polymer residues # ###################### save_chem_comp_DHA _Saveframe_category polymer_residue _Mol_type 'PEPTIDE LINKING' _Name_common '2-AMINO-ACRYLIC ACID' _BMRB_code . _PDB_code DHA _Standard_residue_derivative . _Molecular_mass 87.077 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Fri Feb 3 16:09:25 2012 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? CA CA C . 0 . ? CB CB C . 0 . ? C C C . 0 . ? O O O . 0 . ? OXT OXT O . 0 . ? H H H . 0 . ? H2 H2 H . 0 . ? HB1 HB1 H . 0 . ? HB2 HB2 H . 0 . ? HXT HXT 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 N CA ? ? SING N H ? ? SING N H2 ? ? DOUB CA CB ? ? SING CA C ? ? SING CB HB1 ? ? SING CB HB2 ? ? DOUB C O ? ? SING C OXT ? ? SING OXT HXT ? ? stop_ save_ save_chem_comp_DBU _Saveframe_category polymer_residue _Mol_type 'PEPTIDE LINKING' _Name_common '(2Z)-2-AMINOBUT-2-ENOIC ACID' _BMRB_code . _PDB_code DBU _Standard_residue_derivative . _Molecular_mass 101.104 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Fri Feb 3 16:10:23 2012 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? CA CA C . 0 . ? CB CB C . 0 . ? CG CG C . 0 . ? C C C . 0 . ? O O O . 0 . ? OXT OXT O . 0 . ? H H H . 0 . ? H2 H2 H . 0 . ? HB HB H . 0 . ? HG1 HG1 H . 0 . ? HG2 HG2 H . 0 . ? HG3 HG3 H . 0 . ? HXT HXT 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 N CA ? ? SING N H ? ? SING N H2 ? ? DOUB CA CB ? ? SING CA C ? ? SING CB CG ? ? SING CB HB ? ? SING CG HG1 ? ? SING CG HG2 ? ? SING CG HG3 ? ? DOUB C O ? ? SING C OXT ? ? SING OXT HXT ? ? stop_ save_ save_chem_comp_DAL _Saveframe_category polymer_residue _Mol_type 'D-PEPTIDE LINKING' _Name_common D-ALANINE _BMRB_code . _PDB_code DAL _Standard_residue_derivative . _Molecular_mass 89.093 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Fri Feb 3 16:11:22 2012 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? CA CA C . 0 . ? CB CB C . 0 . ? C C C . 0 . ? O O O . 0 . ? OXT OXT O . 0 . ? H H H . 0 . ? H2 H2 H . 0 . ? HA HA H . 0 . ? HB1 HB1 H . 0 . ? HB2 HB2 H . 0 . ? HB3 HB3 H . 0 . ? HXT HXT 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 N CA ? ? SING N H ? ? SING N H2 ? ? SING CA CB ? ? SING CA C ? ? SING CA HA ? ? SING CB HB1 ? ? SING CB HB2 ? ? SING CB HB3 ? ? DOUB C O ? ? SING C OXT ? ? SING OXT HXT ? ? stop_ save_ save_chem_comp_ABA _Saveframe_category polymer_residue _Mol_type 'L-PEPTIDE LINKING' _Name_common 'ALPHA-AMINOBUTYRIC ACID' _BMRB_code . _PDB_code ABA _Standard_residue_derivative . _Molecular_mass 103.120 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Fri Feb 3 16:12:13 2012 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? CA CA C . 0 . ? C C C . 0 . ? O O O . 0 . ? CB CB C . 0 . ? CG CG C . 0 . ? OXT OXT O . 0 . ? H H H . 0 . ? HN2 HN2 H . 0 . ? HA HA H . 0 . ? HB3 HB3 H . 0 . ? HB2 HB2 H . 0 . ? HG1 HG1 H . 0 . ? HG3 HG3 H . 0 . ? HG2 HG2 H . 0 . ? HXT HXT 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 N CA ? ? SING N H ? ? SING N HN2 ? ? SING CA C ? ? SING CA CB ? ? SING CA HA ? ? DOUB C O ? ? SING C OXT ? ? SING CB CG ? ? SING CB HB3 ? ? SING CB HB2 ? ? SING CG HG1 ? ? SING CG HG3 ? ? SING CG HG2 ? ? SING OXT HXT ? ? stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Cell_line $nisin 'Lactococcus lactis' 1358 Eubacteria . Lactococcus lactis 'NIZO 22186' 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 $nisin 'purified from the natural source' . . . . . 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 $nisin 1.2 mM '[U-100% 15N]' d6-DMSO 99 % . stop_ save_ ############################ # Computer software used # ############################ save_NMRPipe _Saveframe_category software _Name NMRPipe _Version . _Details . save_ save_NMRView _Saveframe_category software _Name NMRView _Version . _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model AVANVE _Field_strength 500 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model AVANVE _Field_strength 600 _Details . save_ save_NMR_spectrometer_3 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model AVANVE _Field_strength 750 _Details . save_ save_NMR_spectrometer_4 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model AVANVE _Field_strength 900 _Details . save_ ############################# # NMR applied experiments # ############################# save_1H-15N_NOESY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N NOESY' _Sample_label $sample_1 save_ save_1H-15N_TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N TOCSY' _Sample_label $sample_1 save_ save_1H-1H_NOESY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H NOESY' _Sample_label $sample_1 save_ save_1H-1H_TOCSY_4 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H TOCSY' _Sample_label $sample_1 save_ save_1H-1H_DQF-COSY_5 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H DQF-COSY' _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_Ex-cond_1 _Saveframe_category sample_conditions _Details 'The sample is disolved in pure DMSO so no pH was measured' loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH . . n/a temperature 300 1 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 DSS C 13 'methyl protons' ppm 0.0 . indirect . . . 0.251449530 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_free_nisin _Saveframe_category assigned_chemical_shifts _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $Ex-cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'nisin monomer' _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 1 ILE CA C 59.855 0.05 1 2 1 1 ILE HA H 3.781 0.03 1 3 1 1 ILE CB C 39.224 0.05 1 4 1 1 ILE HB H 1.872 0.03 1 5 1 1 ILE CG1 C 26.951 0.05 2 6 1 1 ILE HG13 H 1.160 0.03 9 7 1 1 ILE HG12 H 1.533 0.03 9 8 1 1 ILE HD1 H 0.868 0.03 1 9 1 1 ILE HG2 H 0.952 0.03 4 10 1 1 ILE N N 125.066 0.03 1 11 2 2 DBU H1 H 9.738 0.03 1 12 2 2 DBU HB1 H 6.260 0.03 1 13 2 2 DBU CG C 16.239 0.05 1 14 2 2 DBU HG1 H 1.686 0.03 1 15 2 2 DBU N N 113.834 0.03 1 16 3 3 DAL H H 8.144 0.03 1 17 3 3 DAL CA C 57.347 0.05 1 18 3 3 DAL HA H 4.361 0.03 1 19 3 3 DAL CB C 37.374 0.05 1 20 3 3 DAL HB2 H 2.912 0.03 2 21 3 3 DAL HB1 H 3.004 0.03 2 22 4 4 ILE N N 115.251 0.03 1 23 4 4 ILE H H 7.890 0.03 1 24 4 4 ILE CA C 60.976 0.05 1 25 4 4 ILE HA H 4.075 0.03 1 26 4 4 ILE CB C 36.704 0.05 1 27 4 4 ILE HB H 2.001 0.03 1 28 4 4 ILE CG1 C 27.416 0.05 2 29 4 4 ILE HG13 H 1.043 0.03 9 30 4 4 ILE HG12 H 1.352 0.03 9 31 4 4 ILE HD1 H 0.753 0.03 1 32 4 4 ILE HG2 H 0.853 0.03 4 33 5 5 DHA N N 124.873 0.03 1 34 5 5 DHA H H 8.832 0.03 1 35 5 5 DHA CB C 27.064 0.05 1 36 5 5 DHA HB1 H 5.973 0.03 2 37 5 5 DHA HB2 H 5.484 0.03 2 38 6 6 LEU N N 116.587 0.03 1 39 6 6 LEU H H 8.764 0.03 1 40 6 6 LEU CA C 54.856 0.05 1 41 6 6 LEU HA H 4.195 0.03 1 42 6 6 LEU CB C 40.929 0.05 1 43 6 6 LEU HB3 H 1.519 0.03 2 44 6 6 LEU HB2 H 1.669 0.03 2 45 6 6 LEU HG H 1.601 0.03 1 46 6 6 LEU HD1 H 0.851 0.03 4 47 6 6 LEU HD2 H 0.813 0.03 4 48 7 7 CYS N N 117.061 0.03 1 49 7 7 CYS H H 7.950 0.03 1 50 7 7 CYS CA C 57.245 0.05 1 51 7 7 CYS HA H 4.612 0.03 1 52 7 7 CYS CB C 37.736 0.05 1 53 7 7 CYS HB3 H 2.772 0.03 2 54 7 7 CYS HB2 H 3.052 0.03 2 55 8 8 ABA N N 111.053 0.03 1 56 8 8 ABA H H 8.237 0.03 1 57 8 8 ABA CA C 60.041 0.05 1 58 8 8 ABA HA H 4.953 0.03 1 59 8 8 ABA CB C 50.966 0.05 1 60 8 8 ABA HB2 H 3.398 0.03 1 61 8 8 ABA HG1 H 1.152 0.03 1 62 9 9 PRO CA C 66.266 0.05 1 63 9 9 PRO HA H 4.404 0.03 1 64 9 9 PRO CB C 31.698 0.05 1 65 9 9 PRO HB3 H 1.604 0.03 2 66 9 9 PRO HB2 H 2.234 0.03 2 67 9 9 PRO HG3 H 1.749 0.03 2 68 9 9 PRO HG2 H 1.903 0.03 2 69 9 9 PRO CD C 66.221 0.05 1 70 9 9 PRO HD3 H 3.267 0.03 2 71 9 9 PRO HD2 H 3.409 0.03 2 72 10 10 GLY N N 101.541 0.03 1 73 10 10 GLY H H 8.156 0.03 1 74 10 10 GLY CA C 45.003 0.05 1 75 10 10 GLY HA3 H 4.291 0.03 1 76 10 10 GLY HA2 H 4.291 0.03 1 77 11 11 CYS N N 116.935 0.03 1 78 11 11 CYS H H 7.411 0.03 1 79 11 11 CYS CA C 56.008 0.05 1 80 11 11 CYS HA H 3.843 0.03 1 81 11 11 CYS CB C 39.805 0.05 1 82 11 11 CYS HB3 H 2.898 0.03 2 83 11 11 CYS HB2 H 3.430 0.03 2 84 12 12 LYS N N 118.994 0.03 1 85 12 12 LYS H H 8.168 0.03 1 86 12 12 LYS CA C 56.189 0.05 1 87 12 12 LYS HA H 4.271 0.03 1 88 12 12 LYS HB3 H 1.484 0.03 2 89 12 12 LYS HB2 H 1.610 0.03 2 90 12 12 LYS HG3 H 1.237 0.03 2 91 12 12 LYS HG2 H 1.354 0.03 2 92 12 12 LYS HD2 H 1.351 0.03 2 93 12 12 LYS CE C 41.833 0.05 1 94 12 12 LYS HE3 H 2.735 0.03 1 95 12 12 LYS HE2 H 2.735 0.03 1 96 13 13 ABA N N 115.087 0.03 1 97 13 13 ABA H H 7.982 0.03 1 98 13 13 ABA CA C 60.185 0.05 1 99 13 13 ABA HA H 4.399 0.03 1 100 13 13 ABA CB C 46.388 0.05 1 101 13 13 ABA HB2 H 3.263 0.03 1 102 13 13 ABA HG1 H 1.156 0.03 1 103 14 14 GLY N N 108.612 0.03 1 104 14 14 GLY H H 8.199 0.03 1 105 14 14 GLY CA C 45.053 0.05 1 106 14 14 GLY HA3 H 3.852 0.03 1 107 14 14 GLY HA2 H 3.852 0.03 1 108 15 15 ALA N N 123.016 0.03 1 109 15 15 ALA H H 8.509 0.03 1 110 15 15 ALA CA C 53.604 0.05 1 111 15 15 ALA HA H 4.126 0.03 1 112 15 15 ALA HB H 1.257 0.03 1 113 16 16 LEU N N 113.172 0.03 1 114 16 16 LEU H H 8.106 0.03 1 115 16 16 LEU CA C 55.729 0.05 1 116 16 16 LEU HA H 4.060 0.03 1 117 16 16 LEU CB C 42.230 0.05 1 118 16 16 LEU HB3 H 1.478 0.03 2 119 16 16 LEU HB2 H 1.596 0.03 2 120 16 16 LEU HG H 1.260 0.03 1 121 16 16 LEU HD1 H 0.848 0.03 4 122 16 16 LEU HD2 H 0.796 0.03 4 123 17 17 MET N N 112.557 0.03 1 124 17 17 MET H H 7.381 0.03 1 125 17 17 MET CA C 54.593 0.05 1 126 17 17 MET HA H 4.348 0.03 1 127 17 17 MET CB C 32.921 0.05 1 128 17 17 MET HB3 H 2.328 0.03 2 129 17 17 MET HB2 H 2.529 0.03 2 130 17 17 MET CG C 33.780 0.05 1 131 17 17 MET HG3 H 1.979 0.03 2 132 17 17 MET HG2 H 2.015 0.03 2 133 17 17 MET CE C 17.821 0.05 1 134 17 17 MET HE H 1.999 0.03 1 135 18 18 GLY N N 105.006 0.03 1 136 18 18 GLY H H 7.863 0.03 1 137 18 18 GLY CA C 45.383 0.05 1 138 18 18 GLY HA3 H 3.560 0.03 1 139 18 18 GLY HA2 H 3.560 0.03 1 140 19 19 CYS N N 115.300 0.03 1 141 19 19 CYS H H 7.556 0.03 1 142 19 19 CYS CA C 56.829 0.05 1 143 19 19 CYS HA H 4.317 0.03 1 144 19 19 CYS HB3 H 2.683 0.03 2 145 19 19 CYS HB2 H 2.800 0.03 2 146 20 20 ASN N N 117.596 0.03 1 147 20 20 ASN H H 8.254 0.03 1 148 20 20 ASN CA C 53.004 0.05 1 149 20 20 ASN HA H 4.443 0.03 1 150 20 20 ASN CB C 39.991 0.05 1 151 20 20 ASN HB3 H 2.453 0.03 2 152 20 20 ASN HB2 H 2.591 0.03 2 153 20 20 ASN HD21 H 7.458 0.03 2 154 20 20 ASN HD22 H 6.927 0.03 2 155 20 20 ASN ND2 N 110.939 0.03 1 156 21 21 MET CA C 54.533 0.05 1 157 21 21 MET HA H 4.329 0.03 1 158 21 21 MET CB C 32.597 0.05 1 159 21 21 MET HB3 H 2.383 0.03 2 160 21 21 MET HB2 H 2.464 0.03 2 161 21 21 MET CG C 34.262 0.05 1 162 21 21 MET HG3 H 1.761 0.03 2 163 21 21 MET HG2 H 1.952 0.03 2 164 21 21 MET CE C 17.821 0.05 1 165 21 21 MET HE H 1.261 0.03 1 166 22 22 LYS N N 121.123 0.03 1 167 22 22 LYS H H 8.122 0.03 1 168 22 22 LYS CA C 57.044 0.05 1 169 22 22 LYS HA H 4.166 0.03 1 170 22 22 LYS HB3 H 1.536 0.03 2 171 22 22 LYS HB2 H 1.610 0.03 2 172 22 22 LYS HG3 H 1.236 0.03 2 173 22 22 LYS HG2 H 1.352 0.03 2 174 22 22 LYS CD C 25.353 0.05 1 175 22 22 LYS HD2 H 1.251 0.03 2 176 22 22 LYS CE C 41.833 0.05 1 177 22 22 LYS HE3 H 2.727 0.03 1 178 22 22 LYS HE2 H 2.727 0.03 1 179 22 22 LYS HZ H 7.661 0.03 1 180 23 23 ABA N N 113.339 0.03 1 181 23 23 ABA H H 8.399 0.03 1 182 23 23 ABA CA C 60.913 0.05 1 183 23 23 ABA HA H 4.711 0.03 1 184 23 23 ABA CB C 49.394 0.05 1 185 23 23 ABA HB2 H 3.435 0.03 1 186 23 23 ABA HG1 H 1.178 0.03 1 187 24 24 ALA N N 127.087 0.03 1 188 24 24 ALA H H 7.978 0.03 1 189 24 24 ALA CA C 51.851 0.05 1 190 24 24 ALA HA H 4.576 0.03 1 191 24 24 ALA HB H 1.260 0.03 1 192 25 25 ABA N N 115.044 0.03 1 193 25 25 ABA H H 8.705 0.03 1 194 25 25 ABA CA C 61.930 0.05 1 195 25 25 ABA HA H 4.597 0.03 1 196 25 25 ABA CB C 47.979 0.05 1 197 25 25 ABA HB2 H 3.371 0.03 1 198 25 25 ABA HG1 H 1.265 0.03 1 199 26 26 CYS N N 118.633 0.03 1 200 26 26 CYS H H 7.672 0.03 1 201 26 26 CYS CA C 57.910 0.05 1 202 26 26 CYS HA H 3.745 0.03 1 203 26 26 CYS CB C 40.385 0.05 1 204 26 26 CYS HB3 H 2.781 0.03 2 205 26 26 CYS HB2 H 3.405 0.03 2 206 27 27 ASN N N 115.251 0.03 1 207 27 27 ASN H H 7.894 0.03 1 208 27 27 ASN CA C 52.941 0.05 1 209 27 27 ASN HA H 4.512 0.03 1 210 27 27 ASN CB C 38.368 0.05 1 211 27 27 ASN HB3 H 2.447 0.03 2 212 27 27 ASN HB2 H 2.588 0.03 2 213 27 27 ASN HD21 H 7.301 0.03 2 214 27 27 ASN HD22 H 6.778 0.03 2 215 27 27 ASN ND2 N 108.825 0.03 1 216 28 28 CYS N N 115.261 0.03 1 217 28 28 CYS H H 7.355 0.03 1 218 28 28 CYS CA C 55.497 0.05 1 219 28 28 CYS HA H 4.179 0.03 1 220 28 28 CYS CB C 41.586 0.05 1 221 28 28 CYS HB3 H 2.564 0.03 2 222 28 28 CYS HB2 H 3.367 0.03 2 223 29 29 SER N N 113.704 0.03 1 224 29 29 SER H H 8.095 0.03 1 225 29 29 SER CA C 58.026 0.05 1 226 29 29 SER HA H 4.329 0.03 1 227 29 29 SER CB C 64.622 0.05 1 228 29 29 SER HB3 H 3.555 0.03 1 229 29 29 SER HB2 H 3.555 0.03 1 230 30 30 ILE N N 117.041 0.03 1 231 30 30 ILE H H 7.748 0.03 1 232 30 30 ILE CA C 60.181 0.05 1 233 30 30 ILE HA H 4.136 0.03 1 234 30 30 ILE CB C 39.444 0.05 1 235 30 30 ILE HB H 1.689 0.03 1 236 30 30 ILE CG1 C 27.233 0.05 2 237 30 30 ILE HG13 H 1.037 0.03 9 238 30 30 ILE HG12 H 1.284 0.03 9 239 30 30 ILE HD1 H 1.035 0.03 1 240 30 30 ILE HG2 H 0.755 0.03 4 241 31 31 HIS N N 119.607 0.03 1 242 31 31 HIS H H 8.273 0.03 1 243 31 31 HIS CA C 54.417 0.05 1 244 31 31 HIS HA H 4.665 0.03 1 245 31 31 HIS CB C 30.003 0.05 1 246 31 31 HIS HB3 H 2.878 0.03 2 247 31 31 HIS HB2 H 3.059 0.03 2 248 31 31 HIS HE1 H 6.523 0.03 1 249 32 32 VAL N N 115.600 0.03 1 250 32 32 VAL H H 7.879 0.03 1 251 32 32 VAL CA C 61.518 0.05 1 252 32 32 VAL HA H 4.321 0.03 1 253 32 32 VAL HB H 1.995 0.03 1 254 32 32 VAL HG2 H 0.826 0.03 4 255 32 32 VAL HG1 H 0.826 0.03 4 256 33 33 DHA CB C 27.188 0.05 1 257 33 33 DHA HB1 H 6.122 0.03 2 258 33 33 DHA HB2 H 5.556 0.03 2 259 34 34 LYS N N 113.751 0.03 1 260 34 34 LYS H H 8.474 0.03 1 261 34 34 LYS CA C 55.729 0.05 1 262 34 34 LYS HA H 4.067 0.03 1 263 34 34 LYS HB3 H 1.689 0.03 2 264 34 34 LYS HB2 H 1.777 0.03 2 265 34 34 LYS HG3 H 1.247 0.03 2 266 34 34 LYS HG2 H 1.355 0.03 2 267 34 34 LYS HD3 H 1.510 0.03 2 268 34 34 LYS HD2 H 1.595 0.03 2 269 34 34 LYS CE C 41.833 0.05 1 270 34 34 LYS HE3 H 2.751 0.03 1 271 34 34 LYS HE2 H 2.751 0.03 1 stop_ loop_ _Atom_shift_assign_ID_ambiguity 9 32 47,46 122,121 stop_ save_