data_6352 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Isolation and characterization of epilancin 15X, a novel lantibiotic from a clinical strain of Staphylococcus epidermidis ; _BMRB_accession_number 6352 _BMRB_flat_file_name bmr6352.str _Entry_type original _Submission_date 2004-10-14 _Accession_date 2004-10-14 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details . loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Ekkelenkamp Miquel . . 2 Hanssen Micha G.M. . 3 Hsu Shang-Te D. . 4 'de Jong' Ad . . 5 Milatovic Dana . . 6 Verhoef Jan . . 7 'van Nuland' Nico A.J. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 189 "13C chemical shifts" 140 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2008-07-16 update BMRB 'Updating non-standard residue' stop_ _Original_release_date 2004-10-14 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Isolation and structural characterization of epilancin 15X, a novel lantibiotic from a clinical strain of Staphylococcus epidermidis ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 15792796 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Ekkelenkamp Miquel B. . 2 Hanssen Micha G.M. . 3 Hsu Shang-Te D. . 4 'de Jong' Ad . . 5 Milatovic Dana . . 6 Verhoef Jan . . 7 'van Nuland' Nico A.J. . stop_ _Journal_abbreviation 'FEBS Lett.' _Journal_volume 579 _Journal_issue 9 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 1917 _Page_last 1922 _Year 2005 _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. Mapping the targeted membrane pore formation mechanism by solution NMR: the nisin Z and lipid II interaction in SDS micelles. 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_ref_2 _Saveframe_category citation _Citation_full ; Hsu ST, Breukink E, Bierbaum G, Sahl HG, de Kruijff B, Kaptein R, van Nuland NA, Bonvin AM. Free Full Text NMR study of mersacidin and lipid II interaction in dodecylphosphocholine micelles. Conformational changes are a key to antimicrobial activity. J Biol Chem. 2003 Apr 11;278(15):13110-7. Epub 2003 Jan 31. ; _Citation_title ; NMR study of mersacidin and lipid II interaction in dodecylphosphocholine micelles. Conformational changes are a key to antimicrobial activity. ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 12562773 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Hsu 'Shang-Te D.' T. . 2 Breukink Eefjan . . 3 Bierbaum Gabriele . . 4 Sahl Hans-Georg G. . 5 'de Kruijff' Ben . . 6 Kaptein Rob . . 7 'van Nuland' 'Nico A. J.' A. . 8 Bonvin 'Alexandre M. J. J.' M. . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'The Journal of biological chemistry' _Journal_volume 278 _Journal_issue 15 _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 13110 _Page_last 13117 _Year 2003 _Details ; Mersacidin belongs to the type B lantibiotics (lanthionine-containing antibiotics) that contain post-translationally modified amino acids and cyclic ring structures. It targets the cell wall precursor lipid II and thereby inhibits cell wall synthesis. In light of the emerging antibiotics resistance problem, the understanding of the antibacterial activity on a structural basis provides a key to circumvent this issue. Here we present solution NMR studies of mersacidin-lipid II interaction in dodecylphosphocholine (DPC) micelles. Distinct solution structures of mersacidin were determined in three different states: in water/methanol solution and in DPC micelles with and without lipid II. The structures in various sample conditions reveal remarkable conformational changes in which the junction between Ala-12 and Abu-13 (where Abu is aminobutyric acid) effectively serves as the hinge for the opening and closure of the ring structures. The DPC micelle-bound form resembles the previously determined NMR and x-ray crystal structures of mersacidin in pure methanol but substantially deviates from the other two states in our current report. The structural changes delineate the large chemical shift perturbations observed during the course of a two-step (15)N-(1)H heteronuclear single quantum coherence titration. They also modulate the surface charge distribution of mersacidin suggesting that electrostatics play a central role in the mersacidin-lipid II interaction. The observed conformational adaptability of mersacidin might be a general feature of lipid II-interacting antibiotics/peptides. ; save_ save_ref_3 _Saveframe_category citation _Citation_full ; Hsu ST, Breukink E, Tischenko E, Lutters MA, De Kruijff B, Kaptein R, Bonvin AM, Van Nuland NA. The nisin-lipid II complex reveals a pyrophosphate cage that provides a blueprint for novel antibiotics. Nat Struct Mol Biol. 2004 Oct;11(10):963-7. Epub 2004 Sep 12. ; _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.' T. . 2 Breukink Eefjan . . 3 Tischenko Eugene . . 4 Lutters 'Mandy A. G.' A. . 5 'de Kruijff' Ben . . 6 Kaptein Robert . . 7 Bonvin 'Alexandre M. J. J.' M. . 8 'van Nuland' 'Nico A. J.' A. . stop_ _Journal_abbreviation 'Nat. Struct. Mol. Biol.' _Journal_name_full 'Nature structural & molecular biology' _Journal_volume 11 _Journal_issue 10 _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 963 _Page_last 967 _Year 2004 _Details ; The emerging antibiotics-resistance problem has underlined the urgent need for novel antimicrobial agents. Lantibiotics (lanthionine-containing antibiotics) are promising candidates to alleviate this problem. Nisin, a member of this family, has a unique pore-forming activity against bacteria. It binds to lipid II, the essential precursor of cell wall synthesis. As a result, the membrane permeabilization activity of nisin is increased by three orders of magnitude. Here we report the solution structure of the complex of nisin and lipid II. The structure shows a novel lipid II-binding motif in which the pyrophosphate moiety of lipid II is primarily coordinated by the N-terminal backbone amides of nisin via intermolecular hydrogen bonds. This cage structure provides a rationale for the conservation of the lanthionine rings among several lipid II-binding lantibiotics. The structure of the pyrophosphate cage offers a template for structure-based design of novel antibiotics. ; save_ ################################## # Molecular system description # ################################## save_system_Epilancin_15X _Saveframe_category molecular_system _Mol_system_name 'Epilancin 15X' _Abbreviation_common 'Epilancin 15X' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'Epilancin 15X' $Epilancin_15X stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all other bound' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_Epilancin_15X _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'Epilancin 15X' _Abbreviation_common 'Epilancin 15X' _Molecular_mass 3175.2 _Mol_thiol_state 'all other bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 31 _Mol_residue_sequence ; XAXIVKXXIKAXKKLCRGFX LXCGCHFXGKK ; loop_ _Residue_seq_code _Residue_label 1 HOP 2 ALA 3 DHA 4 ILE 5 VAL 6 LYS 7 AA4 8 AA4 9 ILE 10 LYS 11 ALA 12 DAL 13 LYS 14 LYS 15 LEU 16 CYS 17 ARG 18 GLY 19 PHE 20 ABA 21 LEU 22 ABA 23 CYS 24 GLY 25 CYS 26 HIS 27 PHE 28 AA4 29 GLY 30 LYS 31 LYS stop_ _Sequence_homology_query_date 2008-03-24 _Sequence_homology_query_revised_last_date 2007-05-31 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value PDB 1W9N 'A Chain A, Isolation And Characterization OfEpilancin 15x, A Novel Antibiotic From A ClinicalStrain Of Staphylococcus Ep' 100.00 31 100 100 2e-05 stop_ 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 Mon Aug 1 12:11:43 2011 ; 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_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 Mon Aug 1 10:26:43 2011 ; 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_HOP _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common (1S,2S,5S)2-(4-GLUTARIDYLBENZYL)-5-PHENYL-1-CYCLOHEXANOL _BMRB_code . _PDB_code HOP _Standard_residue_derivative . _Molecular_mass 381.465 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Mon Aug 1 12:13:42 2011 ; 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 . ? C6 C6 C . 0 . ? C5 C5 C . 0 . ? C4 C4 C . 0 . ? C3 C3 C . 0 . ? C7 C7 C . 0 . ? C12 C12 C . 0 . ? C11 C11 C . 0 . ? C10 C10 C . 0 . ? C9 C9 C . 0 . ? C8 C8 C . 0 . ? C13 C13 C . 0 . ? C14 C14 C . 0 . ? C15 C15 C . 0 . ? C16 C16 C . 0 . ? C17 C17 C . 0 . ? C18 C18 C . 0 . ? O19 O19 O . 0 . ? C20 C20 C . 0 . ? O21 O21 O . 0 . ? N22 N22 N . 0 . ? C23 C23 C . 0 . ? C24 C24 C . 0 . ? C25 C25 C . 0 . ? C26 C26 C . 0 . ? O27 O27 O . 0 . ? O28 O28 O . 0 . ? H1 H1 H . 0 . ? H2 H2 H . 0 . ? H6 H6 H . 0 . ? H5 H5 H . 0 . ? H3 H3 H . 0 . ? H7 H7 H . 0 . ? H121 H121 H . 0 . ? H122 H122 H . 0 . ? H11 H11 H . 0 . ? H10 H10 H . 0 . ? H91 H91 H . 0 . ? H92 H92 H . 0 . ? H81 H81 H . 0 . ? H82 H82 H . 0 . ? H14 H14 H . 0 . ? H15 H15 H . 0 . ? H17 H17 H . 0 . ? H18 H18 H . 0 . ? HO9 HO9 H . 0 . ? HN2 HN2 H . 0 . ? H231 H231 H . 0 . ? H232 H232 H . 0 . ? H241 H241 H . 0 . ? H242 H242 H . 0 . ? H251 H251 H . 0 . ? H252 H252 H . 0 . ? HO8 HO8 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 C1 C6 ? ? SING C1 H1 ? ? DOUB C2 C3 ? ? SING C2 H2 ? ? SING C6 C5 ? ? SING C6 H6 ? ? DOUB C5 C4 ? ? SING C5 H5 ? ? SING C4 C3 ? ? SING C4 C7 ? ? SING C3 H3 ? ? SING C7 C12 ? ? SING C7 C8 ? ? SING C7 H7 ? ? SING C12 C11 ? ? SING C12 H121 ? ? SING C12 H122 ? ? SING C11 C10 ? ? SING C11 O19 ? ? SING C11 H11 ? ? SING C10 C9 ? ? SING C10 C13 ? ? SING C10 H10 ? ? SING C9 C8 ? ? SING C9 H91 ? ? SING C9 H92 ? ? SING C8 H81 ? ? SING C8 H82 ? ? DOUB C13 C14 ? ? SING C13 C18 ? ? SING C14 C15 ? ? SING C14 H14 ? ? DOUB C15 C16 ? ? SING C15 H15 ? ? SING C16 C17 ? ? SING C16 C20 ? ? DOUB C17 C18 ? ? SING C17 H17 ? ? SING C18 H18 ? ? SING O19 HO9 ? ? DOUB C20 O21 ? ? SING C20 N22 ? ? SING N22 C23 ? ? SING N22 HN2 ? ? SING C23 C24 ? ? SING C23 H231 ? ? SING C23 H232 ? ? SING C24 C25 ? ? SING C24 H241 ? ? SING C24 H242 ? ? SING C25 C26 ? ? SING C25 H251 ? ? SING C25 H252 ? ? DOUB C26 O27 ? ? SING C26 O28 ? ? SING O28 HO8 ? ? stop_ save_ save_chem_comp_AA4 _Saveframe_category polymer_residue _Mol_type 'L-PEPTIDE LINKING' _Name_common '2-AMINO-5-HYDROXYPENTANOIC ACID' _BMRB_code AA4 _PDB_code AA4 _Standard_residue_derivative . _Molecular_mass 133.146 _Mol_paramagnetic . _Details . 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 . ? OE OE 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 . ? HD2 HD2 H . 0 . ? HD3 HD3 H . 0 . ? HE HE 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 ? ? SING CD OE ? ? SING CD HD2 ? ? SING CD HD3 ? ? SING OE HE ? ? 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 ABA _PDB_code ABA _Standard_residue_derivative . _Molecular_mass 103.120 _Mol_paramagnetic . _Details . 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 _Strain $Epilancin_15X 'Staphylococcus epidermidis' 1282 Eubacteria . Staphylococcus epidermidis 15X150 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 $Epilancin_15X 'purified from the natural source' . . . . . stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_water _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $Epilancin_15X 0.8 mM . stop_ save_ save_sample_d2o _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $Epilancin_15X 3.1 mM . stop_ save_ ############################ # Computer software used # ############################ save_NMRPipe _Saveframe_category software _Name NMRPipe _Version n/a loop_ _Task 'data processing' stop_ _Details . save_ save_NMRView _Saveframe_category software _Name NMRView _Version 4.0.5 loop_ _Task Assignment stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DRX _Field_strength 750 _Details . save_ ############################# # NMR applied experiments # ############################# save_1H-1H_TOCSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H TOCSY' _Sample_label . save_ save_1H-1H_NOESY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H NOESY' _Sample_label . save_ save_1H-13C_HSQC_3 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-13C HSQC' _Sample_label . save_ save_1H-13C_HMBC_4 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-13C HMBC' _Sample_label . save_ ####################### # Sample conditions # ####################### save_Ex-cond_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH . . pH temperature 305 . K stop_ save_ save_Ex-cond_2 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH . . pH temperature 283 . 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 _Reference_correction_type water H 1 protons ppm 4.91 internal direct . internal parallel 1.0 temperature 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_Epilancin_15X_283K _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '1H-1H TOCSY' stop_ loop_ _Sample_label $sample_water stop_ _Sample_conditions_label $Ex-cond_2 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'Epilancin 15X' _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 HOP CA C 69.006 0.0 . 2 . 1 HOP HA H 4.257 0.0 . 3 . 1 HOP CB C 21.217 0.0 . 4 . 1 HOP HB1 H 1.327 0.0 . 5 . 1 HOP C C 179.193 0.0 . 6 . 2 ALA H H 8.33 0.0 . 7 . 2 ALA CA C 51.147 0.0 . 8 . 2 ALA HA H 4.31 0.0 . 9 . 2 ALA CB C 17.907 0.0 . 10 . 2 ALA HB H 1.413 0.0 . 11 . 2 ALA C C 175.824 0.0 . 12 . 3 DHA H H 9.892 0.0 . 13 . 3 DHA CA C 137.099 0.0 . 14 . 3 DHA CB C 114.436 0.0 . 15 . 3 DHA HB2 H 5.488 0.0 . 16 . 3 DHA HB3 H 5.562 0.0 . 17 . 3 DHA C C 168.652 0.0 . 18 . 4 ILE H H 8.256 0.0 . 19 . 4 ILE CA C 60.396 0.0 . 20 . 4 ILE HA H 4.161 0.0 . 21 . 4 ILE CB C 37.237 0.0 . 22 . 4 ILE HB H 1.873 0.0 . 23 . 4 ILE CG1 C 26.384 0.0 . 24 . 4 ILE HG13 H 1.46 0.0 . 25 . 4 ILE HG12 H 1.18 0.0 . 26 . 4 ILE CD1 C 11.608 0.0 . 27 . 4 ILE HD1 H 0.846 0.0 . 28 . 4 ILE CG2 C 16.303 0.0 . 29 . 4 ILE HG2 H 0.868 0.0 . 30 . 4 ILE C C 175.344 0.0 . 31 . 5 VAL H H 8.283 0.0 . 32 . 5 VAL CA C 61.151 0.0 . 33 . 5 VAL HA H 4.082 0.0 . 34 . 5 VAL CB C 31.443 0.0 . 35 . 5 VAL HB H 2.019 0.0 . 36 . 5 VAL CG2 C 19.629 0.0 . 37 . 5 VAL HG2 H 0.9 0.0 . 38 . 5 VAL CG1 C 20.037 0.0 . 39 . 5 VAL HG1 H 0.851 0.0 . 40 . 5 VAL C C 175.348 0.0 . 41 . 6 LYS H H 8.516 0.0 . 42 . 6 LYS CA C 55.477 0.0 . 43 . 6 LYS HA H 4.36 0.0 . 44 . 6 LYS CB C 31.735 0.0 . 45 . 6 LYS HB3 H 1.89 0.0 . 46 . 6 LYS HB2 H 1.807 0.0 . 47 . 6 LYS CG C 23.644 0.0 . 48 . 6 LYS HG3 H 1.495 0.0 . 49 . 6 LYS HG2 H 1.415 0.0 . 50 . 6 LYS CD C 27.746 0.0 . 51 . 6 LYS HD3 H 1.67 0.0 . 52 . 6 LYS HD2 H 1.67 0.0 . 53 . 6 LYS CE C 39.455 0.0 . 54 . 6 LYS HE3 H 2.942 0.0 . 55 . 6 LYS HE2 H 2.942 0.0 . 56 . 6 LYS HZ H 7.59 0.0 . 57 . 6 LYS C C 175.457 0.0 . 58 . 7 AA4 H H 9.767 0.0 . 59 . 7 AA4 CA C 129.663 0.0 . 60 . 7 AA4 CB C 135.931 0.0 . 61 . 7 AA4 HB H 6.668 0.0 . 62 . 7 AA4 CG2 C 14.277 0.0 . 63 . 7 AA4 HG21 H 1.781 0.0 . 64 . 7 AA4 C C 167.962 0.0 . 65 . 8 AA4 H H 9.224 0.0 . 66 . 8 AA4 CA C 129.727 0.0 . 67 . 8 AA4 CB C 136.14 0.0 . 68 . 8 AA4 HB H 6.665 0.0 . 69 . 8 AA4 CG2 C 14.283 0.0 . 70 . 8 AA4 HG21 H 1.717 0.0 . 71 . 8 AA4 C C 168.814 0.0 . 72 . 9 ILE H H 7.898 0.0 . 73 . 9 ILE CA C 60.91 0.0 . 74 . 9 ILE HA H 4.103 0.0 . 75 . 9 ILE CB C 37.07 0.0 . 76 . 9 ILE HB H 1.895 0.0 . 77 . 9 ILE CG1 C 26.706 0.0 . 78 . 9 ILE HG13 H 1.456 0.0 . 79 . 9 ILE HG12 H 1.19 0.0 . 80 . 9 ILE CD1 C 11.608 0.0 . 81 . 9 ILE HD1 H 0.821 0.0 . 82 . 9 ILE CG2 C 16.597 0.0 . 83 . 9 ILE HG2 H 0.877 0.0 . 84 . 9 ILE C C 175.903 0.0 . 85 . 10 LYS H H 8.268 0.0 . 86 . 10 LYS CA C 55.526 0.0 . 87 . 10 LYS HA H 4.182 0.0 . 88 . 10 LYS CB C 31.838 0.0 . 89 . 10 LYS HB3 H 1.806 0.0 . 90 . 10 LYS HB2 H 1.755 0.0 . 91 . 10 LYS CG C 23.64 0.0 . 92 . 10 LYS HG3 H 1.456 0.0 . 93 . 10 LYS HG2 H 1.385 0.0 . 94 . 10 LYS CD C 27.844 0.0 . 95 . 10 LYS HD3 H 1.745 0.0 . 96 . 10 LYS HD2 H 1.745 0.0 . 97 . 10 LYS CE C 40.755 0.0 . 98 . 10 LYS HE3 H 2.943 0.0 . 99 . 10 LYS HE2 H 2.943 0.0 . 100 . 10 LYS HZ H 7.57 0.0 . 101 . 11 ALA H H 8.034 0.0 . 102 . 11 ALA CA C 51.525 0.0 . 103 . 11 ALA HA H 4.218 0.0 . 104 . 11 ALA CB C 17.946 0.0 . 105 . 11 ALA HB H 1.364 0.0 . 106 . 11 ALA C C 176.776 0.0 . 107 . 12 DAL H H 8.061 0.0 . 108 . 12 DAL CA C 55.762 0.0 . 109 . 12 DAL HA H 4.401 0.0 . 110 . 12 DAL CB C 35.694 0.0 . 111 . 12 DAL HB1 H 3.07 0.0 . 112 . 12 DAL HB2 H 3.025 0.0 . 113 . 12 DAL C C 179.654 0.0 . 114 . 13 LYS H H 8.392 0.0 . 115 . 13 LYS CA C 55.087 0.0 . 116 . 13 LYS HA H 4.104 0.0 . 117 . 13 LYS CB C 20.996 0.0 . 118 . 13 LYS HB3 H 1.766 0.0 . 119 . 13 LYS HB2 H 1.766 0.0 . 120 . 13 LYS HG3 H 1.395 0.0 . 121 . 13 LYS HG2 H 1.245 0.0 . 122 . 13 LYS HD3 H 1.632 0.0 . 123 . 13 LYS HD2 H 1.597 0.0 . 124 . 13 LYS HE3 H 2.957 0.0 . 125 . 13 LYS HE2 H 2.957 0.0 . 126 . 13 LYS HZ H 7.577 0.0 . 127 . 13 LYS C C 180.353 0.0 . 128 . 14 LYS H H 7.801 0.0 . 129 . 14 LYS CA C 55.814 0.0 . 130 . 14 LYS HA H 3.994 0.0 . 131 . 14 LYS CB C 31.723 0.0 . 132 . 14 LYS HB3 H 1.763 0.0 . 133 . 14 LYS HB2 H 1.763 0.0 . 134 . 14 LYS CG C 23.644 0.0 . 135 . 14 LYS HG3 H 1.315 0.0 . 136 . 14 LYS HG2 H 1.315 0.0 . 137 . 14 LYS CD C 27.746 0.0 . 138 . 14 LYS HD3 H 1.64 0.0 . 139 . 14 LYS HD2 H 1.64 0.0 . 140 . 14 LYS CE C 40.639 0.0 . 141 . 14 LYS HE3 H 2.943 0.0 . 142 . 14 LYS HE2 H 2.943 0.0 . 143 . 14 LYS HZ H 7.581 0.0 . 144 . 14 LYS C C 175.46 0.0 . 145 . 15 LEU H H 8.386 0.0 . 146 . 15 LEU CA C 54.283 0.0 . 147 . 15 LEU HA H 4.207 0.0 . 148 . 15 LEU CB C 39.55 0.0 . 149 . 15 LEU HB3 H 1.671 0.0 . 150 . 15 LEU HB2 H 1.626 0.0 . 151 . 15 LEU CG C 26.053 0.0 . 152 . 15 LEU HG H 1.538 0.0 . 153 . 15 LEU CD1 C 23.801 0.0 . 154 . 15 LEU HD1 H 0.809 0.0 . 155 . 15 LEU CD2 C 21.768 0.0 . 156 . 15 LEU HD2 H 0.858 0.0 . 157 . 15 LEU C C 177.654 0.0 . 158 . 16 CYS H H 8.018 0.0 . 159 . 16 CYS CA C 54.641 0.0 . 160 . 16 CYS HA H 4.477 0.0 . 161 . 16 CYS CB C 27.465 0.0 . 162 . 16 CYS HB3 H 3.067 0.0 . 163 . 16 CYS HB2 H 3.015 0.0 . 164 . 16 CYS C C 173.209 0.0 . 165 . 17 ARG H H 8.396 0.0 . 166 . 17 ARG CA C 55.605 0.0 . 167 . 17 ARG HA H 4.241 0.0 . 168 . 17 ARG CB C 29.122 0.0 . 169 . 17 ARG HB3 H 1.83 0.0 . 170 . 17 ARG HB2 H 1.775 0.0 . 171 . 17 ARG CG C 25.941 0.0 . 172 . 17 ARG HG3 H 1.644 0.0 . 173 . 17 ARG HG2 H 1.578 0.0 . 174 . 17 ARG CD C 42.045 0.0 . 175 . 17 ARG HD3 H 3.167 0.0 . 176 . 17 ARG HD2 H 3.167 0.0 . 177 . 17 ARG HE H 7.216 0.0 . 178 . 17 ARG CZ C 158.259 0.0 . 179 . 17 ARG HH21 H 6.913 0.0 . 180 . 17 ARG HH22 H 6.913 0.0 . 181 . 17 ARG HH11 H 6.483 0.0 . 182 . 17 ARG HH12 H 6.483 0.0 . 183 . 17 ARG C C 178.051 0.0 . 184 . 18 GLY H H 8.66 0.0 . 185 . 18 GLY CA C 44.029 0.0 . 186 . 18 GLY HA3 H 3.983 0.0 . 187 . 18 GLY HA2 H 3.819 0.0 . 188 . 18 GLY C C 173.444 0.0 . 189 . 19 PHE H H 8.046 0.0 . 190 . 19 PHE CA C 57.711 0.0 . 191 . 19 PHE HA H 4.561 0.0 . 192 . 19 PHE CB C 37.919 0.0 . 193 . 19 PHE HB3 H 3.196 0.0 . 194 . 19 PHE HB2 H 2.938 0.0 . 195 . 19 PHE CG C 137.172 0.0 . 196 . 19 PHE CD1 C 130.694 0.0 . 197 . 19 PHE HD1 H 7.213 0.0 . 198 . 19 PHE CE1 C 130.482 0.0 . 199 . 19 PHE HE1 H 7.316 0.0 . 200 . 19 PHE CZ C 128.973 0.0 . 201 . 19 PHE HZ H 7.256 0.0 . 202 . 19 PHE CE2 C 130.482 0.0 . 203 . 19 PHE HE2 H 7.316 0.0 . 204 . 19 PHE CD2 C 130.694 0.0 . 205 . 19 PHE HD2 H 7.213 0.0 . 206 . 19 PHE C C 177.07 0.0 . 207 . 20 ABA H H 8.758 0.0 . 208 . 20 ABA CA C 60.175 0.0 . 209 . 20 ABA HA H 4.685 0.0 . 210 . 20 ABA CB C 49.041 0.0 . 211 . 20 ABA HB H 3.296 0.0 . 212 . 20 ABA CG2 C 21.092 0.0 . 213 . 20 ABA HG2 H 0.55 0.0 . 214 . 20 ABA C C 176.762 0.0 . 215 . 21 LEU H H 7.991 0.0 . 216 . 21 LEU CA C 54.461 0.0 . 217 . 21 LEU HA H 4.56 0.0 . 218 . 21 LEU CB C 39.081 0.0 . 219 . 21 LEU HB3 H 1.771 0.0 . 220 . 21 LEU HB2 H 1.53 0.0 . 221 . 21 LEU CG C 26.185 0.0 . 222 . 21 LEU HG H 1.413 0.0 . 223 . 21 LEU CD1 C 22.479 0.0 . 224 . 21 LEU HD1 H 0.873 0.0 . 225 . 21 LEU CD2 C 22.719 0.0 . 226 . 21 LEU HD2 H 0.92 0.0 . 227 . 21 LEU C C 177.632 0.0 . 228 . 22 ABA H H 9.581 0.0 . 229 . 22 ABA CA C 60.932 0.0 . 230 . 22 ABA HA H 4.87 0.0 . 231 . 22 ABA CB C 46.744 0.0 . 232 . 22 ABA HB H 3.44 0.0 . 233 . 22 ABA CG2 C 21.173 0.0 . 234 . 22 ABA HG2 H 1.344 0.0 . 235 . 23 CYS H H 7.692 0.0 . 236 . 23 CYS CA C 57.21 0.0 . 237 . 23 CYS HA H 3.991 0.0 . 238 . 23 CYS CB C 37.945 0.0 . 239 . 23 CYS HB3 H 3.516 0.0 . 240 . 23 CYS HB2 H 2.736 0.0 . 241 . 23 CYS C C 172.718 0.0 . 242 . 24 GLY H H 9.118 0.0 . 243 . 24 GLY CA C 44.539 0.0 . 244 . 24 GLY HA3 H 3.989 0.0 . 245 . 24 GLY HA2 H 3.735 0.0 . 246 . 24 GLY C C 172.557 0.0 . 247 . 25 CYS H H 7.333 0.0 . 248 . 25 CYS CA C 54.744 0.0 . 249 . 25 CYS HA H 3.937 0.0 . 250 . 25 CYS CB C 38.533 0.0 . 251 . 25 CYS HB3 H 3.361 0.0 . 252 . 25 CYS HB2 H 2.499 0.0 . 253 . 25 CYS C C 173.156 0.0 . 254 . 26 HIS H H 8.734 0.0 . 255 . 26 HIS CA C 53.61 0.0 . 256 . 26 HIS HA H 4.695 0.0 . 257 . 26 HIS CB C 27.59 0.0 . 258 . 26 HIS HB3 H 3.189 0.0 . 259 . 26 HIS HB2 H 3.065 0.0 . 260 . 26 HIS CG C 129.924 0.0 . 261 . 26 HIS CD2 C 118.772 0.0 . 262 . 26 HIS HD2 H 7.203 0.0 . 263 . 26 HIS CE1 C 135.03 0.0 . 264 . 26 HIS HE1 H 8.572 0.0 . 265 . 26 HIS C C 173.053 0.0 . 266 . 27 PHE H H 8.605 0.0 . 267 . 27 PHE CA C 56.915 0.0 . 268 . 27 PHE HA H 4.676 0.0 . 269 . 27 PHE CB C 38.164 0.0 . 270 . 27 PHE HB3 H 3.112 0.0 . 271 . 27 PHE HB2 H 3.112 0.0 . 272 . 27 PHE CG C 137.403 0.0 . 273 . 27 PHE CD1 C 130.738 0.0 . 274 . 27 PHE HD1 H 7.275 0.0 . 275 . 27 PHE CE1 C 130.643 0.0 . 276 . 27 PHE HE1 H 7.351 0.0 . 277 . 27 PHE CZ C 128.959 0.0 . 278 . 27 PHE HZ H 7.294 0.0 . 279 . 27 PHE CE2 C 130.643 0.0 . 280 . 27 PHE HE2 H 7.351 0.0 . 281 . 27 PHE CD2 C 130.738 0.0 . 282 . 27 PHE HD2 H 7.275 0.0 . 283 . 27 PHE C C 174.791 0.0 . 284 . 28 AA4 H H 9.684 0.0 . 285 . 28 AA4 CA C 128.979 0.0 . 286 . 28 AA4 CB C 136.488 0.0 . 287 . 28 AA4 HB H 6.623 0.0 . 288 . 28 AA4 CG2 C 14.129 0.0 . 289 . 28 AA4 HG21 H 1.426 0.0 . 290 . 28 AA4 C C 168.367 0.0 . 291 . 29 GLY H H 7.829 0.0 . 292 . 29 GLY CA C 44.078 0.0 . 293 . 29 GLY HA3 H 3.891 0.0 . 294 . 29 GLY HA2 H 3.891 0.0 . 295 . 29 GLY C C 172.915 0.0 . 296 . 30 LYS H H 8.006 0.0 . 297 . 30 LYS CA C 55.007 0.0 . 298 . 30 LYS HA H 4.279 0.0 . 299 . 30 LYS CB C 32.055 0.0 . 300 . 30 LYS HB3 H 1.832 0.0 . 301 . 30 LYS HB2 H 1.752 0.0 . 302 . 30 LYS CG C 23.64 0.0 . 303 . 30 LYS HG3 H 1.419 0.0 . 304 . 30 LYS HG2 H 1.419 0.0 . 305 . 30 LYS CD C 27.746 0.0 . 306 . 30 LYS HD3 H 1.642 0.0 . 307 . 30 LYS HD2 H 1.642 0.0 . 308 . 30 LYS CE C 40.639 0.0 . 309 . 30 LYS HE3 H 2.94 0.0 . 310 . 30 LYS HE2 H 2.94 0.0 . 311 . 30 LYS HZ H 7.536 0.0 . 312 . 30 LYS C C 175.163 0.0 . 313 . 31 LYS H H 8.208 0.0 . 314 . 31 LYS CA C 54.87 0.0 . 315 . 31 LYS HA H 4.1 0.0 . 316 . 31 LYS CB C 31.842 0.0 . 317 . 31 LYS HB3 H 1.778 0.0 . 318 . 31 LYS HB2 H 1.682 0.0 . 319 . 31 LYS CG C 23.644 0.0 . 320 . 31 LYS HG3 H 1.371 0.0 . 321 . 31 LYS HG2 H 1.371 0.0 . 322 . 31 LYS CD C 27.746 0.0 . 323 . 31 LYS HD3 H 1.619 0.0 . 324 . 31 LYS HD2 H 1.619 0.0 . 325 . 31 LYS CE C 40.639 0.0 . 326 . 31 LYS HE3 H 2.941 0.0 . 327 . 31 LYS HE2 H 2.941 0.0 . 328 . 31 LYS HZ H 7.514 0.0 . 329 . 31 LYS C C 176.375 0.0 . stop_ save_