data_6146 ####################### # 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 6146 _BMRB_flat_file_name bmr6146.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 2 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 236 "13C chemical shifts" 73 "15N chemical shifts" 34 "31P chemical shifts" 2 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2008-07-16 update BMRB 'Updating non-standard residue' stop_ loop_ _Related_BMRB_accession_number _Relationship 6144 'nisin monomer' 6145 '3LII in DMSO' 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_references _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_ save_references_1 _Saveframe_category citation _Citation_full ; Breukink E, van Heusden HE, Vollmerhaus PJ, Swiezewska E, Brunner L, Walker S, Heck AJ, de Kruijff B. J Biol Chem. 2003 May 30;278(22):19898-903. ; _Citation_title 'Lipid II is an intrinsic component of the pore induced by nisin in bacterial membranes.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 12663672 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Breukink Eefjan . . 2 'van Heusden' 'Hester E' E. . 3 Vollmerhaus 'Pauline J' J. . 4 Swiezewska Ewa . . 5 Brunner Livia . . 6 Walker Suzanne . . 7 Heck 'Albert J R' J. . 8 'de Kruijff' Ben . . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'The Journal of biological chemistry' _Journal_volume 278 _Journal_issue 22 _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 19898 _Page_last 19903 _Year 2003 _Details ; The peptidoglycan layers surrounding bacterial membranes are essential for bacterial cell survival and provide an important target for antibiotics. Many antibiotics have mechanisms of action that involve binding to Lipid II, the prenyl chain-linked donor of the peptidoglycan building blocks. One of these antibiotics, the pore-forming peptide nisin uses Lipid II as a receptor molecule to increase its antimicrobial efficacy dramatically. Nisin is the first example of a targeted membrane-permeabilizing peptide antibiotic. However, it was not known whether Lipid II functions only as a receptor to recruit nisin to bacterial membranes, thus increasing its specificity for bacterial cells, or whether it also plays a role in pore formation. We have developed a new method to produce large amounts of Lipid II and variants thereof so that we can address the role of the lipid-linked disaccharide in the activity of nisin. We show here that Lipid II is not only the receptor for nisin but an intrinsic component of the pore formed by nisin, and we present a new model for the pore complex that includes Lipid II. ; save_ ################################## # Molecular system description # ################################## save_nisin_3LII_complex _Saveframe_category molecular_system _Mol_system_name 'nisin/3LII complex' _Abbreviation_common 'nisin/3LII complex' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'nisin monomer' $nisin 3LII $3LII 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 DHB_2 3 3 ABA_2 4 4 ILE 5 5 DHA_2 6 6 LEU 7 7 CYS 8 8 MLN_2 9 9 PRO 10 10 GLY 11 11 CYS 12 12 LYS 13 13 MLN_2 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 MLN_2 24 24 ALA 25 25 MLN_2 26 26 CYS 27 27 ASN 28 28 CYS 29 29 SER 30 30 ILE 31 31 HIS 32 32 VAL 33 33 DHA_2 34 34 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ save_3LII _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'G-gamma-glutamic acid' _Molecular_mass . _Mol_thiol_state 'not present' _Details . _Residue_count 11 _Mol_residue_sequence AXKXXXXXXXX loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 1 ALA 2 2 DGL 3 3 LYS 4 4 DAL 5 5 DAL 6 6 NAG 7 7 NAM_2 8 8 POP 9 9 TPL_2 10 10 TPL_2 11 11 TPL_2 stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ ###################### # Polymer residues # ###################### save_DHA_2 _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common dehydroalanine _Abbreviation_common DHA_2 _BMRB_code DHA_2 _PDB_code DHA_2 _Standard_residue_derivative SER loop_ _Mol_label _Residue_seq_code $nisin 5 $nisin 33 stop_ _Molecular_mass . _Mol_paramagnetic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N . N . 0 . ? CA . C . 0 . ? CB . C . 0 . ? C . C . 0 . ? O . O . 0 . ? H . H . 0 . ? HB2 . H . 0 . ? HB3 . 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 ? ? DOUB CA CB ? ? SING CA C ? ? SING CB HB2 ? ? SING CB HB3 ? ? DOUB C O ? ? stop_ save_ save_DHB_2 _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common dehydrobutyrine _Abbreviation_common DHB_2 _BMRB_code DHB_2 _PDB_code . _Standard_residue_derivative . loop_ _Mol_label _Residue_seq_code $nisin 2 stop_ _Molecular_mass . _Mol_paramagnetic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N . N . 0 . ? CA . C . 0 . ? CB . C . 0 . ? CG . C . 0 . ? C . C . 0 . ? O . O . 0 . ? H . H . 0 . ? HB2 . H . 0 . ? HG1 . H . 0 . ? HG2 . H . 0 . ? HG3 . 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 ? ? DOUB CA CB ? ? SING CA C ? ? SING CB HB2 ? ? SING CB CG ? ? SING CG HG1 ? ? SING CG HG2 ? ? SING CG HG3 ? ? DOUB C O ? ? stop_ save_ save_ABA_2 _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common 'lanthionine or meso-lanthionine D-alanine moiety' _Abbreviation_common ABA_2 _BMRB_code ABA_2 _PDB_code . _Standard_residue_derivative SER loop_ _Mol_label _Residue_seq_code $nisin 3 stop_ _Molecular_mass . _Mol_paramagnetic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N . N . 0 . ? CA . C . 0 . ? CB . C . 0 . ? CG . C . 0 . ? C . C . 0 . ? O . O . 0 . ? H . H . 0 . ? HB2 . H . 0 . ? HB3 . 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 CA CB ? ? SING CA HA ? ? SING CA C ? ? SING CB HB2 ? ? SING CB HB3 ? ? DOUB C O ? ? stop_ save_ save_MLN_2 _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common ''3-methyllanthionine or (2S, 3S, 6R)-3-methyllanthionine' _Abbreviation_common MLN_2 _BMRB_code MLN_2 _PDB_code . _Standard_residue_derivative THR loop_ _Mol_label _Residue_seq_code $nisin 8 $nisin 13 $nisin 23 $nisin 25 stop_ _Molecular_mass . _Mol_paramagnetic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N . N . 0 . ? CA . C . 0 . ? CB . C . 0 . ? CG . C . 0 . ? C . C . 0 . ? O . O . 0 . ? H . H . 0 . ? HA . H . 0 . ? HB2 . H . 0 . ? HG1 . H . 0 . ? HG2 . H . 0 . ? HB3 . 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 CA CB ? ? SING CA HA ? ? SING CA C ? ? SING CB HB2 ? ? SING CB CG ? ? SING CG HG1 ? ? SING CG HG2 ? ? SING CG HG3 ? ? DOUB C O ? ? stop_ save_ save_chem_comp_DGL _Saveframe_category polymer_residue _Mol_type 'D-PEPTIDE LINKING' _Name_common 'D-GLUTAMIC ACID' _BMRB_code . _PDB_code DGL _Standard_residue_derivative . _Molecular_mass 147.129 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Thu Feb 9 15:26:35 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 . ? CD CD C . 0 . ? OE1 OE1 O . 0 . ? OE2 OE2 O . 0 . ? OXT OXT O . 0 . ? H H H . 0 . ? H2 H2 H . 0 . ? HA HA H . 0 . ? HB2 HB2 H . 0 . ? HB3 HB3 H . 0 . ? HG2 HG2 H . 0 . ? HG3 HG3 H . 0 . ? HE2 HE2 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 C ? ? SING CA CB ? ? SING CA HA ? ? DOUB C O ? ? SING C OXT ? ? SING CB CG ? ? SING CB HB2 ? ? SING CB HB3 ? ? SING CG CD ? ? SING CG HG2 ? ? SING CG HG3 ? ? DOUB CD OE1 ? ? SING CD OE2 ? ? SING OE2 HE2 ? ? 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 Thu Feb 9 15:27:57 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_NAG _Saveframe_category polymer_residue _Mol_type D-SACCHARIDE _Name_common N-ACETYL-D-GLUCOSAMINE _BMRB_code . _PDB_code NAG _Standard_residue_derivative . _Molecular_mass 221.208 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Thu Feb 9 15:29:52 2012 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons C1 C1 C . 0 . ? C2 C2 C . 0 . ? C3 C3 C . 0 . ? C4 C4 C . 0 . ? C5 C5 C . 0 . ? C6 C6 C . 0 . ? C7 C7 C . 0 . ? C8 C8 C . 0 . ? N2 N2 N . 0 . ? O1 O1 O . 0 . ? O3 O3 O . 0 . ? O4 O4 O . 0 . ? O5 O5 O . 0 . ? O6 O6 O . 0 . ? O7 O7 O . 0 . ? H1 H1 H . 0 . ? H2 H2 H . 0 . ? H3 H3 H . 0 . ? H4 H4 H . 0 . ? H5 H5 H . 0 . ? H61 H61 H . 0 . ? H62 H62 H . 0 . ? H81 H81 H . 0 . ? H82 H82 H . 0 . ? H83 H83 H . 0 . ? HN2 HN2 H . 0 . ? HO1 HO1 H . 0 . ? HO3 HO3 H . 0 . ? HO4 HO4 H . 0 . ? HO6 HO6 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 C1 C2 ? ? SING C1 O1 ? ? SING C1 O5 ? ? SING C1 H1 ? ? SING C2 C3 ? ? SING C2 N2 ? ? SING C2 H2 ? ? SING C3 C4 ? ? SING C3 O3 ? ? SING C3 H3 ? ? SING C4 C5 ? ? SING C4 O4 ? ? SING C4 H4 ? ? SING C5 C6 ? ? SING C5 O5 ? ? SING C5 H5 ? ? SING C6 O6 ? ? SING C6 H61 ? ? SING C6 H62 ? ? SING C7 C8 ? ? SING C7 N2 ? ? DOUB C7 O7 ? ? SING C8 H81 ? ? SING C8 H82 ? ? SING C8 H83 ? ? SING N2 HN2 ? ? SING O1 HO1 ? ? SING O3 HO3 ? ? SING O4 HO4 ? ? SING O6 HO6 ? ? stop_ save_ save_NAM_2 _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common 'N-acetyl muramic acid' _Abbreviation_common MurNAc _BMRB_code NAM_2 _PDB_code . _Standard_residue_derivative . loop_ _Mol_label _Residue_seq_code $3LII 7 stop_ _Molecular_mass . _Mol_paramagnetic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons C1 . C . 0 . ? C2 . C . 0 . ? C3 . C . 0 . ? C4 . C . 0 . ? C5 . C . 0 . ? C6 . C . 0 . ? C7 . C . 0 . ? C8 . C . 0 . ? C9 . C . 0 . ? C10 . C . 0 . ? C11 . C . 0 . ? N2 . N . 0 . ? O1 . O . 0 . ? O3 . O . 0 . ? O4 . O . 0 . ? O5 . O . 0 . ? O6 . O . 0 . ? O7 . O . 0 . ? O10 . O . 0 . ? H1 . H . 0 . ? HN2 . H . 0 . ? H3 . H . 0 . ? H4 . H . 0 . ? H5 . H . 0 . ? H61 . H . 0 . ? H62 . H . 0 . ? HO6 . H . 0 . ? H81 . H . 0 . ? H82 . H . 0 . ? H83 . H . 0 . ? H9 . H . 0 . ? H111 . H . 0 . ? H112 . H . 0 . ? H113 . 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 C1 C2 ? ? SING C1 O5 ? ? SING C1 O1 ? ? SING C1 H1 ? ? SING C2 C3 ? ? SING C2 N2 ? ? SING N2 HN2 ? ? SING N2 C7 ? ? DOUB C7 O7 ? ? SING C7 C8 ? ? SING C8 H81 ? ? SING C8 H82 ? ? SING C8 H83 ? ? SING C3 C4 ? ? SING C3 O3 ? ? SING C3 H3 ? ? SING O3 C9 ? ? SING C9 H9 ? ? SING C9 C10 ? ? DOUB C10 O10 ? ? SING C9 C11 ? ? SING C11 H111 ? ? SING C11 H112 ? ? SING C11 H113 ? ? SING C4 C5 ? ? SING C4 H4 ? ? SING C4 O4 ? ? SING C5 C6 ? ? SING C5 O5 ? ? SING C5 H5 ? ? SING C6 O6 ? ? SING C6 H61 ? ? SING C6 H62 ? ? SING O6 HO6 ? ? stop_ save_ save_chem_comp_POP _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common 'PYROPHOSPHATE 2-' _BMRB_code . _PDB_code POP _Standard_residue_derivative . _Molecular_mass 175.959 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Thu Feb 9 15:33:42 2012 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons P1 P1 P . 0 . ? O1 O1 O . 0 . ? O2 O2 O . 0 . ? O3 O3 O . -1 . ? O O O . 0 . ? P2 P2 P . 0 . ? O4 O4 O . 0 . ? O5 O5 O . 0 . ? O6 O6 O . -1 . ? HO2 HO2 H . 0 . ? HO5 HO5 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 DOUB P1 O1 ? ? SING P1 O2 ? ? SING P1 O3 ? ? SING P1 O ? ? SING O2 HO2 ? ? SING O P2 ? ? DOUB P2 O4 ? ? SING P2 O5 ? ? SING P2 O6 ? ? SING O5 HO5 ? ? stop_ save_ save_TPL_2 _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common 1,4-trans-isoprene _Abbreviation_common TPL_2 _BMRB_code TPL_2 _PDB_code . _Standard_residue_derivative . loop_ _Mol_label _Residue_seq_code $3LII 9 $3LII 10 $3LII 11 stop_ _Molecular_mass . _Mol_paramagnetic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons C1 . C . 0 . ? C2 . C . 0 . ? C3 . C . 0 . ? C4 . C . 0 . ? C5 . C . 0 . ? H11 . H . 0 . ? H12 . H . 0 . ? H2 . H . 0 . ? H41 . H . 0 . ? H42 . H . 0 . ? H51 . H . 0 . ? H52 . H . 0 . ? H53 . 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 C1 C2 ? ? DOUB C2 C3 ? ? SING C3 C4 ? ? SING C3 C5 ? ? SING C1 H11 ? ? SING C1 H12 ? ? SING C2 H2 ? ? SING C3 C4 ? ? SING C3 C5 ? ? SING C4 H41 ? ? SING C4 H42 ? ? SING C5 H51 ? ? SING C5 H52 ? ? SING C5 H53 ? ? stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $3LII 'Lactococcus lactis' 1358 Eubacteria . Lactococcus lactis $nisin 'Lactococcus lactis' 1358 Eubacteria . Lactococcus lactis 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 $3LII 'enzymatic semisynthesis' . . . . . $nisin 'enzymatic semisynthesis' . . . . . 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 $3LII 1.2 mM . $nisin 1.2 mM '[U-100% 15N]' d6-DMSO 99.9 % . 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 AVANCE _Field_strength 500 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model AVANCE _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_ save_1H-13C_HSQC_6 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-13C HSQC' _Sample_label $sample_1 save_ save_1H-13C_HMBC_7 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-13C HMBC' _Sample_label $sample_1 save_ save_31P_direct_detection_8 _Saveframe_category NMR_applied_experiment _Experiment_name '31P direct detection' _Sample_label $sample_1 save_ save_1H-31P_HSQC_9 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-31P HSQC' _Sample_label $sample_1 save_ save_31P-edited_1H-15N_CT_HSQC_10 _Saveframe_category NMR_applied_experiment _Experiment_name '31P-edited 1H-15N CT HSQC' _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 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_chemical_shift_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 HA H 4.303 0.03 1 2 1 1 ILE CB C 36.014 0.05 1 3 1 1 ILE HB H 2.006 0.03 1 4 1 1 ILE CG1 C 27.053 0.05 2 5 1 1 ILE HG13 H 0.975 0.03 1 6 1 1 ILE HG12 H 0.975 0.03 1 7 1 1 ILE HD1 H 0.825 0.03 1 8 1 1 ILE HG2 H 0.764 0.03 1 9 2 2 DHB_2 N N 139.087 0.03 1 10 2 2 DHB_2 H H 12.663 0.03 1 11 2 2 DHB_2 HB H 6.033 0.03 1 12 2 2 DHB_2 CG2 C 15.423 0.05 1 13 2 2 DHB_2 HG21 H 1.642 0.03 1 14 3 3 ABA_2 N N 112.563 0.03 1 15 3 3 ABA_2 H H 7.867 0.03 1 16 3 3 ABA_2 CA C 56.408 0.05 1 17 3 3 ABA_2 HA H 4.276 0.03 1 18 3 3 ABA_2 HB2 H 3.066 0.03 2 19 3 3 ABA_2 HB1 H 3.250 0.03 2 20 4 4 ILE N N 115.293 0.03 1 21 4 4 ILE H H 8.327 0.03 1 22 4 4 ILE CA C 62.453 0.05 1 23 4 4 ILE HA H 3.967 0.03 1 24 4 4 ILE CB C 38.830 0.05 1 25 4 4 ILE HB H 1.782 0.03 1 26 4 4 ILE CG1 C 27.869 0.05 2 27 4 4 ILE HG13 H 1.107 0.03 2 28 4 4 ILE HG12 H 1.362 0.03 2 29 4 4 ILE CD1 C 13.992 0.05 1 30 4 4 ILE HD1 H 0.655 0.03 1 31 4 4 ILE HG2 H 0.778 0.03 4 32 5 5 DHA_2 N N 123.031 0.03 1 33 5 5 DHA_2 H H 8.224 0.03 1 34 5 5 DHA_2 HB1 H 6.253 0.03 2 35 5 5 DHA_2 HB2 H 5.458 0.03 2 36 6 6 LEU N N 118.175 0.03 1 37 6 6 LEU H H 9.380 0.03 1 38 6 6 LEU CA C 55.685 0.05 1 39 6 6 LEU HA H 3.568 0.03 1 40 6 6 LEU HB3 H 1.602 0.03 2 41 6 6 LEU HB2 H 1.539 0.03 2 42 6 6 LEU HG H 1.694 0.03 1 43 6 6 LEU HD1 H 0.832 0.03 4 44 6 6 LEU HD2 H 0.808 0.03 4 45 7 7 CYS N N 116.767 0.03 1 46 7 7 CYS H H 8.516 0.03 1 47 7 7 CYS CA C 55.167 0.05 1 48 7 7 CYS HA H 4.987 0.03 1 49 7 7 CYS CB C 27.801 0.05 1 50 7 7 CYS HB3 H 2.322 0.03 2 51 7 7 CYS HB2 H 3.053 0.03 2 52 8 8 MLN_2 N N 113.599 0.03 1 53 8 8 MLN_2 H H 10.209 0.03 1 54 8 8 MLN_2 CA C 59.984 0.05 1 55 8 8 MLN_2 HA H 4.788 0.03 1 56 8 8 MLN_2 CB C 50.304 0.05 1 57 8 8 MLN_2 HB H 3.300 0.03 1 58 8 8 MLN_2 HG21 H 1.031 0.03 1 59 9 9 PRO CA C 66.159 0.05 1 60 9 9 PRO HA H 4.409 0.03 1 61 9 9 PRO HB3 H 2.165 0.03 2 62 9 9 PRO HB2 H 1.449 0.03 2 63 9 9 PRO CG C 33.785 0.05 1 64 9 9 PRO HG3 H 1.710 0.03 2 65 9 9 PRO HG2 H 1.756 0.03 2 66 9 9 PRO HD3 H 3.187 0.03 2 67 9 9 PRO HD2 H 3.624 0.03 2 68 10 10 GLY N N 104.715 0.03 1 69 10 10 GLY H H 9.231 0.03 1 70 10 10 GLY HA3 H 4.196 0.03 2 71 10 10 GLY HA2 H 4.238 0.03 2 72 11 11 CYS N N 116.421 0.03 1 73 11 11 CYS H H 7.007 0.03 1 74 11 11 CYS CA C 55.244 0.05 1 75 11 11 CYS HA H 3.781 0.03 1 76 11 11 CYS HB3 H 2.911 0.03 1 77 11 11 CYS HB2 H 2.911 0.03 1 78 12 12 LYS N N 119.608 0.03 1 79 12 12 LYS H H 8.570 0.03 1 80 12 12 LYS HA H 4.103 0.03 1 81 12 12 LYS HB3 H 1.494 0.03 1 82 12 12 LYS HB2 H 1.494 0.03 1 83 12 12 LYS HG3 H 1.249 0.03 1 84 12 12 LYS HG2 H 1.249 0.03 1 85 12 12 LYS HD2 H 1.296 0.03 2 86 12 12 LYS HE3 H 2.719 0.03 1 87 12 12 LYS HE2 H 2.719 0.03 1 88 13 13 MLN_2 N N 113.433 0.03 1 89 13 13 MLN_2 H H 7.865 0.03 1 90 13 13 MLN_2 CA C 60.417 0.05 1 91 13 13 MLN_2 HA H 4.326 0.03 1 92 13 13 MLN_2 CB C 46.419 0.05 1 93 13 13 MLN_2 HB H 3.335 0.03 1 94 13 13 MLN_2 HG21 H 1.136 0.03 1 95 14 14 GLY N N 108.487 0.03 1 96 14 14 GLY H H 8.166 0.03 1 97 14 14 GLY CA C 44.833 0.05 1 98 14 14 GLY HA3 H 3.871 0.03 1 99 14 14 GLY HA2 H 3.871 0.03 1 100 15 15 ALA N N 118.614 0.03 1 101 15 15 ALA H H 8.644 0.03 1 102 15 15 ALA HA H 4.410 0.03 1 103 16 16 LEU N N 112.869 0.03 1 104 16 16 LEU H H 8.370 0.03 1 105 16 16 LEU CA C 55.904 0.05 1 106 16 16 LEU HA H 4.021 0.03 1 107 16 16 LEU HB3 H 1.595 0.03 1 108 16 16 LEU HB2 H 1.595 0.03 1 109 16 16 LEU HG H 1.252 0.03 1 110 16 16 LEU HD1 H 0.770 0.03 4 111 16 16 LEU HD2 H 0.759 0.03 4 112 17 17 MET N N 112.758 0.03 1 113 17 17 MET H H 7.243 0.03 1 114 17 17 MET HA H 4.306 0.03 1 115 17 17 MET CB C 33.486 0.05 1 116 17 17 MET HB3 H 2.062 0.03 1 117 17 17 MET HB2 H 2.062 0.03 1 118 17 17 MET HG3 H 2.057 0.03 2 119 17 17 MET HG2 H 2.080 0.03 2 120 18 18 GLY N N 103.008 0.03 1 121 18 18 GLY H H 7.576 0.03 1 122 18 18 GLY HA3 H 3.549 0.03 1 123 18 18 GLY HA2 H 3.549 0.03 1 124 19 19 CYS N N 114.361 0.03 1 125 19 19 CYS H H 8.167 0.03 1 126 19 19 CYS CA C 56.907 0.05 1 127 19 19 CYS HA H 4.216 0.03 1 128 19 19 CYS HB3 H 2.591 0.03 2 129 19 19 CYS HB2 H 2.795 0.03 2 130 20 20 ASN N N 116.361 0.03 1 131 20 20 ASN H H 8.364 0.03 1 132 20 20 ASN CA C 53.078 0.05 1 133 20 20 ASN HA H 4.413 0.03 1 134 20 20 ASN CB C 39.794 0.05 1 135 20 20 ASN HB3 H 2.474 0.03 2 136 20 20 ASN HB2 H 2.575 0.03 2 137 20 20 ASN ND2 N 111.061 0.03 1 138 20 20 ASN HD21 H 7.439 0.03 2 139 20 20 ASN HD22 H 6.932 0.03 2 140 21 21 MET N N 117.410 0.03 1 141 21 21 MET H H 7.913 0.03 1 142 21 21 MET CA C 54.499 0.05 1 143 21 21 MET HA H 4.313 0.03 1 144 21 21 MET CB C 32.513 0.05 1 145 21 21 MET HB3 H 1.755 0.03 2 146 21 21 MET HB2 H 2.368 0.03 2 147 21 21 MET CG C 34.272 0.05 1 148 21 21 MET HG3 H 1.923 0.03 2 149 21 21 MET HG2 H 2.453 0.03 2 150 22 22 LYS N N 121.123 0.03 1 151 22 22 LYS H H 8.169 0.03 1 152 22 22 LYS HA H 4.148 0.03 1 153 22 22 LYS HB3 H 1.579 0.03 1 154 22 22 LYS HB2 H 1.579 0.03 1 155 22 22 LYS HG3 H 1.252 0.03 1 156 22 22 LYS HG2 H 1.252 0.03 1 157 22 22 LYS HD2 H 1.347 0.03 2 158 22 22 LYS HE3 H 2.734 0.03 1 159 22 22 LYS HE2 H 2.734 0.03 1 160 23 23 MLN_2 N N 113.339 0.03 1 161 23 23 MLN_2 H H 8.399 0.03 1 162 23 23 MLN_2 CA C 60.891 0.05 1 163 23 23 MLN_2 HA H 4.706 0.03 1 164 23 23 MLN_2 CB C 49.604 0.05 1 165 23 23 MLN_2 HB H 3.408 0.03 1 166 23 23 MLN_2 HG21 H 1.162 0.03 1 167 24 24 ALA N N 127.281 0.03 1 168 24 24 ALA H H 7.968 0.03 1 169 24 24 ALA CA C 51.897 0.05 1 170 24 24 ALA HA H 4.563 0.03 1 171 24 24 ALA CB C 20.138 0.05 1 172 24 24 ALA HB H 1.252 0.03 1 173 25 25 MLN_2 N N 115.216 0.03 1 174 25 25 MLN_2 H H 8.681 0.03 1 175 25 25 MLN_2 CA C 61.971 0.05 1 176 25 25 MLN_2 HA H 4.568 0.03 1 177 25 25 MLN_2 CB C 47.840 0.05 1 178 25 25 MLN_2 HB H 3.345 0.03 1 179 25 25 MLN_2 HG21 H 1.247 0.03 1 180 26 26 CYS N N 118.666 0.03 1 181 26 26 CYS H H 7.647 0.03 1 182 26 26 CYS CA C 58.002 0.05 1 183 26 26 CYS HA H 3.702 0.03 1 184 26 26 CYS CB C 40.740 0.05 1 185 26 26 CYS HB3 H 2.749 0.03 2 186 26 26 CYS HB2 H 3.384 0.03 2 187 27 27 ASN N N 115.228 0.03 1 188 27 27 ASN H H 7.862 0.03 1 189 27 27 ASN CA C 52.968 0.05 1 190 27 27 ASN HA H 4.486 0.03 1 191 27 27 ASN CB C 38.360 0.05 1 192 27 27 ASN HB3 H 2.434 0.03 2 193 27 27 ASN HB2 H 2.559 0.03 2 194 27 27 ASN ND2 N 108.783 0.03 1 195 27 27 ASN HD21 H 7.283 0.03 2 196 27 27 ASN HD22 H 6.748 0.03 2 197 28 28 CYS N N 115.457 0.03 1 198 28 28 CYS H H 7.379 0.03 1 199 28 28 CYS CA C 55.253 0.05 1 200 28 28 CYS HA H 4.197 0.03 1 201 28 28 CYS HB3 H 2.550 0.03 2 202 28 28 CYS HB2 H 3.346 0.03 2 203 29 29 SER N N 114.560 0.03 1 204 29 29 SER H H 8.205 0.03 1 205 29 29 SER CA C 58.221 0.05 1 206 29 29 SER HA H 4.295 0.03 1 207 29 29 SER CB C 64.527 0.05 1 208 29 29 SER HB3 H 3.554 0.03 1 209 29 29 SER HB2 H 3.554 0.03 1 210 30 30 ILE N N 116.527 0.03 1 211 30 30 ILE H H 7.710 0.03 1 212 30 30 ILE HA H 4.122 0.03 1 213 30 30 ILE HB H 1.673 0.03 1 214 30 30 ILE CG1 C 27.057 0.05 2 215 30 30 ILE HG13 H 1.004 0.03 1 216 30 30 ILE HG12 H 1.250 0.03 1 217 30 30 ILE HD1 H 1.001 0.03 1 218 30 30 ILE CG2 C 14.420 0.05 2 219 30 30 ILE HG2 H 0.727 0.03 1 220 31 31 HIS N N 120.301 0.03 1 221 31 31 HIS H H 8.077 0.03 1 222 31 31 HIS CA C 55.727 0.05 1 223 31 31 HIS HA H 4.508 0.03 1 224 31 31 HIS CB C 32.403 0.05 1 225 31 31 HIS HB3 H 2.753 0.03 2 226 31 31 HIS HB2 H 2.909 0.03 2 227 31 31 HIS HD2 H 6.725 0.03 2 228 32 32 VAL N N 115.253 0.03 1 229 32 32 VAL H H 7.938 0.03 1 230 32 32 VAL CA C 61.786 0.05 1 231 32 32 VAL HA H 4.174 0.03 1 232 32 32 VAL HB H 1.999 0.03 1 233 32 32 VAL HG2 H 0.801 0.03 4 234 32 32 VAL HG1 H 0.809 0.03 4 235 33 33 DHA_2 N N 126.596 0.03 1 236 33 33 DHA_2 H H 9.169 0.03 1 237 33 33 DHA_2 HB1 H 5.958 0.03 2 238 33 33 DHA_2 HB2 H 5.499 0.03 2 239 34 34 LYS N N 117.620 0.03 1 240 34 34 LYS H H 8.053 0.03 1 241 34 34 LYS CA C 56.467 0.05 1 242 34 34 LYS HA H 4.053 0.03 1 243 34 34 LYS HB3 H 1.635 0.03 2 244 34 34 LYS HB2 H 1.722 0.03 2 245 34 34 LYS HG3 H 1.285 0.03 1 246 34 34 LYS HG2 H 1.285 0.03 1 247 34 34 LYS CE C 41.822 0.05 1 248 34 34 LYS HE3 H 2.724 0.03 1 249 34 34 LYS HE2 H 2.724 0.03 1 stop_ loop_ _Atom_shift_assign_ID_ambiguity 31 '31,31' '44,44,44,43,43,43' '111,111,111,110,110,110' '234,234,234,233,233,233' stop_ save_ save_chemical_shift_3LII _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 3LII _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 ALA H H 7.114 0.03 1 2 1 1 ALA CA C 58.208 0.03 1 3 1 1 ALA HA H 4.112 0.03 1 4 1 1 ALA CB C 19.484 0.03 1 5 1 1 ALA HB H 1.229 0.03 1 6 2 2 DGL H H 7.302 0.03 1 7 2 2 DGL CA C 56.160 0.03 1 8 2 2 DGL HA H 3.824 0.03 1 9 2 2 DGL CB C 5.681 0.03 1 10 2 2 DGL HB2 H 1.659 0.03 2 11 2 2 DGL HB1 H 1.659 0.03 2 12 2 2 DGL CG C 9.307 0.03 1 13 2 2 DGL HG2 H 1.981 0.03 2 14 2 2 DGL HG1 H 2.105 0.03 2 15 3 3 LYS H H 7.951 0.03 1 16 3 3 LYS CA C 56.023 0.03 1 17 3 3 LYS HA H 4.141 0.03 1 18 3 3 LYS HB3 H 1.548 0.03 2 19 3 3 LYS HB2 H 1.617 0.03 2 20 3 3 LYS HG3 H 1.321 0.03 2 21 3 3 LYS HG2 H 1.374 0.03 2 22 3 3 LYS HD2 H 1.486 0.03 2 23 3 3 LYS HE3 H 2.754 0.03 1 24 3 3 LYS HE2 H 2.754 0.03 1 25 4 4 DAL H H 8.047 0.03 1 26 4 4 DAL CA C 51.769 0.03 1 27 4 4 DAL HA H 4.121 0.03 1 28 4 4 DAL CB C 21.205 0.03 1 29 4 4 DAL HB1 H 1.168 0.03 1 30 5 5 DAL H H 7.530 0.03 1 31 5 5 DAL CA C 52.022 0.03 1 32 5 5 DAL HA H 3.909 0.03 1 33 5 5 DAL CB C 20.810 0.03 1 34 5 5 DAL HB1 H 1.178 0.03 1 35 6 6 NAG H1 H 4.279 0.03 1 36 6 6 NAG C2 C 67.971 0.03 1 37 6 6 NAG H2 H 3.602 0.03 1 38 6 6 NAG HN2 H 8.110 0.03 1 39 6 6 NAG H3 H 3.249 0.03 1 40 6 6 NAG C4 C 73.846 0.03 1 41 6 6 NAG H4 H 3.085 0.03 1 42 6 6 NAG C5 C 79.377 0.03 1 43 6 6 NAG H5 H 3.005 0.03 1 44 6 6 NAG H61 H 3.251 0.03 9 45 6 6 NAG H62 H 3.069 0.03 9 46 6 6 NAG C8 C 25.830 0.03 1 47 6 6 NAG H81 H 1.989 0.03 4 48 6 6 NAG H82 H 1.989 0.03 4 49 6 6 NAG H83 H 1.989 0.03 4 50 7 7 NAM_2 H1 H 4.931 0.03 1 51 7 7 NAM_2 C2 C 56.135 0.03 1 52 7 7 NAM_2 H2 H 3.862 0.03 1 53 7 7 NAM_2 HN2 H 7.021 0.03 1 54 7 7 NAM_2 C3 C 75.881 0.03 1 55 7 7 NAM_2 H3 H 3.613 0.03 1 56 7 7 NAM_2 C4 C 75.416 0.03 1 57 7 7 NAM_2 H4 H 3.714 0.03 1 58 7 7 NAM_2 C8 C 26.135 0.03 1 59 7 7 NAM_2 H81 H 1.813 0.03 4 60 7 7 NAM_2 H82 H 1.813 0.03 4 61 7 7 NAM_2 H83 H 1.813 0.03 4 62 7 7 NAM_2 C9 C 79.997 0.03 1 63 7 7 NAM_2 H9 H 4.133 0.03 1 64 7 7 NAM_2 C11 C 22.326 0.03 2 65 7 7 NAM_2 H111 H 1.297 0.03 4 66 7 7 NAM_2 H112 H 1.297 0.03 4 67 7 7 NAM_2 H113 H 1.297 0.03 4 68 8 8 POP PA P -14.140 . 1 69 8 8 POP PB P -13.550 . 1 70 9 9 TPL_2 C1 C 64.601 0.03 1 71 9 9 TPL_2 H11 H 4.328 0.03 9 72 9 9 TPL_2 H12 H 4.243 0.03 9 73 9 9 TPL_2 H2 H 5.654 0.03 1 74 9 9 TPL_2 H41 H 1.891 0.03 9 75 9 9 TPL_2 H42 H 1.538 0.03 9 76 9 9 TPL_2 C5 C 18.937 0.03 1 77 9 9 TPL_2 H51 H 1.606 0.03 4 78 9 9 TPL_2 H52 H 1.606 0.03 4 79 9 9 TPL_2 H53 H 1.606 0.03 4 80 10 10 TPL_2 C1 C 29.212 0.03 1 81 10 10 TPL_2 H11 H 2.097 0.03 9 82 10 10 TPL_2 H12 H 1.988 0.03 9 83 10 10 TPL_2 H2 H 5.062 0.03 1 84 10 10 TPL_2 H41 H 1.592 0.03 9 85 10 10 TPL_2 C5 C 18.898 0.03 1 86 10 10 TPL_2 H51 H 1.537 0.03 4 87 10 10 TPL_2 H52 H 1.537 0.03 4 88 10 10 TPL_2 H53 H 1.537 0.03 4 89 11 11 TPL_2 H2 H 5.009 0.03 1 90 11 11 TPL_2 C4 C 28.608 0.03 1 91 11 11 TPL_2 H41 H 1.591 0.03 9 92 11 11 TPL_2 H42 H 1.591 0.03 9 93 11 11 TPL_2 C5 C 20.656 0.03 1 94 11 11 TPL_2 H51 H 1.517 0.03 4 95 11 11 TPL_2 H52 H 1.517 0.03 4 96 11 11 TPL_2 H53 H 1.517 0.03 4 stop_ loop_ _Atom_shift_assign_ID_ambiguity 49 '48,47' '65,61,59' '67,66' '79,78,77' '88,87,86' '96,95,94' stop_ save_