data_6192 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H, 13C, and 15N Chemical Shift Assignments for Omega-atracotoxin-Hv1a at pH 3.6 ; _BMRB_accession_number 6192 _BMRB_flat_file_name bmr6192.str _Entry_type original _Submission_date 2004-04-23 _Accession_date 2004-04-27 _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 Tedford Hugo W. . 2 Maciejewski Mark W. . 3 King Glenn F. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 203 "13C chemical shifts" 123 "15N chemical shifts" 39 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2004-06-30 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 4233 'H chemical shift by a different group' 5702 'chemical shifts at pH 6.0' 6194 'chemical shifts at pH 6.0' stop_ _Original_release_date 2004-04-27 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; 1H, 13C, and 15N Chemical Shift Assignments for Omega-atracotoxin-Hv1a at pH 3.6 ; _Citation_status 'in preparation' _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Tedford Hugo W. . 2 Maciejewski Mark W. . 3 King Glenn F. . stop_ _Journal_abbreviation . _Journal_volume . _Journal_issue . _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first . _Page_last . _Year . _Details . loop_ _Keyword 'calcium channel' 'cystine knot' 'insecticidal toxin' neurotoxin omega-atracotoxin-1 stop_ save_ ####################################### # Cited references within the entry # ####################################### save_Ref_1 _Saveframe_category citation _Citation_full ; Fletcher JI, Smith R, O'Donoghue SI, Nilges M, Connor M, Howden ME, Christie MJ, King GF. The structure of a novel insecticidal neurotoxin, omega-atracotoxin-HV1, from the venom of an Australian funnel web spider. Nat Struct Biol. 1997 Jul;4(7):559-66. ; _Citation_title ; The structure of a novel insecticidal neurotoxin, omega-atracotoxin-HV1, from the venom of an Australian funnel web spider. ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 9228949 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Fletcher 'J I' I. . 2 Smith R . . 3 O'Donoghue 'S I' I. . 4 Nilges M . . 5 Connor M . . 6 Howden 'M E' E. . 7 Christie 'M J' J. . 8 King 'G F' F. . stop_ _Journal_abbreviation 'Nat. Struct. Biol.' _Journal_name_full 'Nature structural biology' _Journal_volume 4 _Journal_issue 7 _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 559 _Page_last 566 _Year 1997 _Details ; A family of potent insecticidal toxins has recently been isolated from the venom of Australian funnel web spiders. Among these is the 37-residue peptide omega-atracotoxin-HV1 (omega-ACTX-HV1) from Hadronyche versuta. We have chemically synthesized and folded omega-ACTX-HV1, shown that it is neurotoxic, ascertained its disulphide bonding pattern, and determined its three-dimensional solution structure using NMR spectroscopy. The structure consists of a solvent-accessible beta-hairpin protruding from a disulphide-bonded globular core comprising four beta-turns. The three intramolecular disulphide bonds from a cystine knot motif similar to that seen in several other neurotoxic peptides. Despite limited sequence identity, omega-ACTX-HV1 displays significant structural homology with the omega-agatoxins and omega-conotoxins, both of which are vertebrate calcium channel antagonists; however, in contrast with these toxins, we show that omega-ACTX-HV1 inhibits insect, but not mammalian, voltage-gated calcium channel currents. ; save_ save_Ref_2 _Saveframe_category citation _Citation_full ; Tedford HW, Fletcher JI, King GF. Functional significance of the beta hairpin in the insecticidal neurotoxin omega-atracotoxin-Hv1a. J Biol Chem. 2001 Jul 13;276(28):26568-76. ; _Citation_title ; Functional significance of the beta hairpin in the insecticidal neurotoxin omega-atracotoxin-Hv1a. ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11313356 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Tedford 'H W' W. . 2 Fletcher 'J I' I. . 3 King 'G F' F. . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'The Journal of biological chemistry' _Journal_volume 276 _Journal_issue 28 _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 26568 _Page_last 26576 _Year 2001 _Details ; omega-Atracotoxin-Hv1a is an insect-specific neurotoxin whose phylogenetic specificity derives from its ability to antagonize insect, but not vertebrate, voltage-gated calcium channels. In order to help understand its mechanism of action and to enhance its utility as a lead compound for insecticide development, we used a combination of protein engineering and site-directed mutagenesis to probe the toxin for key functional regions. First, we constructed a Hairpinless mutant in which the C-terminal beta-hairpin, which is highly conserved in this family of neurotoxins, was excised without affecting the fold of the residual disulfide-rich core of the toxin. The Hairpinless mutant was devoid of insecticidal activity, indicating the functional importance of the hairpin. We subsequently developed a highly efficient system for production of recombinant toxin and then probed the hairpin for key functional residues using alanine-scanning mutagenesis followed by a second round of mutagenesis based on initial "hits" from the alanine scan. This revealed that two spatially proximal residues, Asn(27) and Arg(35), form a contiguous molecular surface that is essential for toxin activity. We propose that this surface of the beta-hairpin is a key site for interaction of the toxin with insect calcium channels. ; save_ ################################## # Molecular system description # ################################## save_system_Omega-ACTX-Hv1a _Saveframe_category molecular_system _Mol_system_name Omega-atracotoxin-Hv1a _Abbreviation_common Omega-ACTX-Hv1a _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label Omega-ACTX-Hv1a $Omega-ACTX-Hv1a stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all disulfide bound' loop_ _Biological_function 'insect calcium channel blocker' 'insecticidal neurotoxin' stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_Omega-ACTX-Hv1a _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common Omega-atracotoxin-Hv1a _Abbreviation_common Omega-ACTX-Hv1a _Molecular_mass . _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 37 _Mol_residue_sequence ; SPTCIPSGQPCPYNENCCSQ SCTFKENENGNTVKRCD ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 -1 SER 2 1 PRO 3 2 THR 4 3 CYS 5 4 ILE 6 5 PRO 7 6 SER 8 7 GLY 9 8 GLN 10 9 PRO 11 10 CYS 12 11 PRO 13 12 TYR 14 13 ASN 15 14 GLU 16 15 ASN 17 16 CYS 18 17 CYS 19 18 SER 20 19 GLN 21 20 SER 22 21 CYS 23 22 THR 24 23 PHE 25 24 LYS 26 25 GLU 27 26 ASN 28 27 GLU 29 28 ASN 30 29 GLY 31 30 ASN 32 31 THR 33 32 VAL 34 33 LYS 35 34 ARG 36 35 CYS 37 36 ASP stop_ _Sequence_homology_query_date 2008-08-19 _Sequence_homology_query_revised_last_date 2008-08-19 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 BMRB 4233 omega-atracotoxin-Hv1a 100.00 37 100.00 100.00 1.34e-12 BMRB 4937 'Hairpinless mutant of omega-atracotoxin-Hv1a' 54.05 25 100.00 100.00 1.29e-02 BMRB 5702 Omega-atracotoxin-Hv1a 100.00 37 100.00 100.00 1.34e-12 BMRB 6194 Omega-atracotoxin-Hv1a 100.00 37 100.00 100.00 1.34e-12 PDB 1AXH 'Atracotoxin-Hvi From Hadronyche Versuta (Australian Funnel- Web Spider, Nmr, 20 Structures' 100.00 37 100.00 100.00 1.34e-12 PDB 1HVW 'Hairpinless Mutant Of Omega-Atracotoxin-Hv1a' 54.05 25 100.00 100.00 1.29e-02 EMBL CAI79354 'omega atracotoxin [synthetic construct]' 100.00 38 100.00 100.00 1.15e-12 GenBank ABP63656 'omega-atracotoxin-Ar1d [Atrax robustus]' 100.00 78 100.00 100.00 8.27e-14 SWISS-PROT P56207 'Omega-atracotoxin-Hv1a (Omega-AcTx-Hv1a) (AcTx-Hv1)' 100.00 37 100.00 100.00 1.34e-12 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Organ $Omega-ACTX-Hv1a 'Blue Mountains Funnel-Web Spider' 6904 Eukaryota Metazoa Hadronyche versuta 'venom gland' 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 _Details $Omega-ACTX-Hv1a 'recombinant technology' . . . . . ; GST-fusion expression system as described by Tedford et al. (2001) J. Biol. Chem. 276:26568-26576. ; 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 _Concentration_min_value _Concentration_max_value _Isotopic_labeling $Omega-ACTX-Hv1a . mM 2 5 '[U-13C; U-15N]' NaCl 50 mM . . . 'sodium phosphate' 20 mM . . . stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $Omega-ACTX-Hv1a . mM 2 5 '[U-13C; U-15N]' D2O 100 % . . . stop_ save_ ############################ # Computer software used # ############################ save_nmrPipe _Saveframe_category software _Name nmrPipe _Version . loop_ _Task 'spectral processing' stop_ _Details . save_ save_Xeasy _Saveframe_category software _Name Xeasy _Version . loop_ _Task 'resonance assignment' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model INOVA _Field_strength 500 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model INOVA _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_3D_1H-15N_NOESY-HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-15N NOESY-HSQC' _Sample_label . save_ save_3D_1H-13C_HSQC-NOESY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-13C HSQC-NOESY' _Sample_label . save_ save_2D_HNCO_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D HNCO' _Sample_label . save_ save_2D_HNCACO_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D HNCACO' _Sample_label . save_ save_3D_(H)CCOHN-TOCSY_5 _Saveframe_category NMR_applied_experiment _Experiment_name '3D (H)CCOHN-TOCSY' _Sample_label . save_ save_3D_1H-15N_TOCSY_6 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-15N TOCSY' _Sample_label . save_ save_3D_HNHA_7 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNHA' _Sample_label . save_ save_3D_HNHB_8 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNHB' _Sample_label . save_ save_2D_1H-1H_NOESY_9 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _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 'ionic strength' 0.010 0.002 M pH 3.6 0.1 pH temperature 298 1 K stop_ save_ save_Ex-cond_2 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 0.010 0.002 M pH* 3.6 0.1 pH temperature 298 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 C 13 'methyl protons' ppm 0.0 . indirect . . . 0.251449530 DSS H 1 'methyl protons' ppm 0.0 internal direct . . . 1.0 DSS N 15 'methyl protons' ppm 0.0 . indirect . . . 0.101329118 stop_ save_ ################################### # Assigned chemical shift lists # ################################### ################################################################### # Chemical Shift Ambiguity Index Value Definitions # # # # The values other than 1 are used for those atoms with different # # chemical shifts that cannot be assigned to stereospecific atoms # # or to specific residues or chains. # # # # Index Value Definition # # # # 1 Unique (including isolated methyl protons, # # geminal atoms, and geminal methyl # # groups with identical chemical shifts) # # (e.g. ILE HD11, HD12, HD13 protons) # # 2 Ambiguity of geminal atoms or geminal methyl # # proton groups (e.g. ASP HB2 and HB3 # # protons, LEU CD1 and CD2 carbons, or # # LEU HD11, HD12, HD13 and HD21, HD22, # # HD23 methyl protons) # # 3 Aromatic atoms on opposite sides of # # symmetrical rings (e.g. TYR HE1 and HE2 # # protons) # # 4 Intraresidue ambiguities (e.g. LYS HG and # # HD protons or TRP HZ2 and HZ3 protons) # # 5 Interresidue ambiguities (LYS 12 vs. LYS 27) # # 6 Intermolecular ambiguities (e.g. ASP 31 CA # # in monomer 1 and ASP 31 CA in monomer 2 # # of an asymmetrical homodimer, duplex # # DNA assignments, or other assignments # # that may apply to atoms in one or more # # molecule in the molecular assembly) # # 9 Ambiguous, specific ambiguity not defined # # # ################################################################### save_CSR_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '3D 1H-15N NOESY-HSQC' '3D 1H-13C HSQC-NOESY' '2D HNCO' '2D HNCACO' '3D (H)CCOHN-TOCSY' '3D 1H-15N TOCSY' '3D HNHA' '3D HNHB' '2D 1H-1H NOESY' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $Ex-cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name Omega-ACTX-Hv1a _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 2 PRO CD C 50.7 0.3 1 2 1 2 PRO CA C 63.1 0.3 1 3 1 2 PRO HA H 4.56 0.02 1 4 1 2 PRO CB C 32.2 0.3 1 5 1 2 PRO HB2 H 2.02 0.02 2 6 1 2 PRO HB3 H 2.36 0.02 2 7 1 2 PRO CG C 27.3 0.3 1 8 1 2 PRO HG2 H 2.06 0.02 1 9 1 2 PRO HG3 H 2.06 0.02 1 10 1 2 PRO HD2 H 3.77 0.02 2 11 1 2 PRO HD3 H 3.87 0.02 2 12 1 2 PRO C C 176.6 0.3 1 13 2 3 THR N N 113.8 0.2 1 14 2 3 THR H H 8.29 0.02 1 15 2 3 THR CA C 61.3 0.3 1 16 2 3 THR HA H 4.41 0.02 1 17 2 3 THR CB C 69.7 0.3 1 18 2 3 THR HB H 4.29 0.02 1 19 2 3 THR HG2 H 1.24 0.02 1 20 2 3 THR CG2 C 21.6 0.3 1 21 2 3 THR C C 174.0 0.3 1 22 3 4 CYS N N 118.9 0.2 1 23 3 4 CYS H H 8.02 0.02 1 24 3 4 CYS CA C 54.1 0.3 1 25 3 4 CYS HA H 4.85 0.02 1 26 3 4 CYS CB C 42.6 0.3 1 27 3 4 CYS HB2 H 3.00 0.02 2 28 3 4 CYS HB3 H 3.21 0.02 2 29 3 4 CYS C C 173.4 0.3 1 30 4 5 ILE N N 122.7 0.2 1 31 4 5 ILE H H 9.18 0.02 1 32 4 5 ILE CA C 59.8 0.3 1 33 4 5 ILE HA H 4.42 0.02 1 34 4 5 ILE CB C 39.9 0.3 1 35 4 5 ILE HB H 1.91 0.02 1 36 4 5 ILE HG2 H 1.16 0.02 1 37 4 5 ILE CG2 C 19.0 0.3 1 38 4 5 ILE CG1 C 27.3 0.3 1 39 4 5 ILE HG12 H 1.13 0.02 2 40 4 5 ILE HG13 H 1.78 0.02 2 41 4 5 ILE HD1 H 1.00 0.02 1 42 4 5 ILE CD1 C 14.1 0.3 1 43 5 6 PRO CD C 52.2 0.3 1 44 5 6 PRO CA C 62.4 0.3 1 45 5 6 PRO HA H 4.48 0.02 1 46 5 6 PRO CB C 33.4 0.3 1 47 5 6 PRO HB2 H 1.96 0.02 2 48 5 6 PRO HB3 H 2.42 0.02 2 49 5 6 PRO CG C 27.4 0.3 1 50 5 6 PRO HG2 H 1.95 0.02 2 51 5 6 PRO HG3 H 2.04 0.02 2 52 5 6 PRO HD2 H 3.64 0.02 2 53 5 6 PRO HD3 H 4.23 0.02 2 54 5 6 PRO C C 175.4 0.3 1 55 6 7 SER N N 115.2 0.2 1 56 6 7 SER H H 8.45 0.02 1 57 6 7 SER CA C 60.8 0.3 1 58 6 7 SER HA H 3.84 0.02 1 59 6 7 SER CB C 62.9 0.3 1 60 6 7 SER HB2 H 3.67 0.02 1 61 6 7 SER HB3 H 3.67 0.02 1 62 6 7 SER C C 175.2 0.3 1 63 7 8 GLY N N 112.7 0.2 1 64 7 8 GLY H H 9.45 0.02 1 65 7 8 GLY CA C 44.8 0.3 1 66 7 8 GLY HA2 H 3.67 0.02 2 67 7 8 GLY HA3 H 4.40 0.02 2 68 7 8 GLY C C 174.2 0.3 1 69 8 9 GLN N N 119.4 0.2 1 70 8 9 GLN H H 7.39 0.02 1 71 8 9 GLN CA C 53.3 0.3 1 72 8 9 GLN HA H 4.74 0.02 1 73 8 9 GLN CB C 27.9 0.3 1 74 8 9 GLN HB2 H 2.02 0.02 2 75 8 9 GLN HB3 H 2.16 0.02 2 76 8 9 GLN CG C 33.7 0.3 1 77 8 9 GLN HG2 H 2.24 0.02 2 78 8 9 GLN HG3 H 2.37 0.02 2 79 8 9 GLN NE2 N 113.4 0.2 1 80 8 9 GLN HE21 H 6.93 0.02 2 81 8 9 GLN HE22 H 7.66 0.02 2 82 9 10 PRO CD C 51.0 0.3 1 83 9 10 PRO CA C 63.2 0.3 1 84 9 10 PRO HA H 4.96 0.02 1 85 9 10 PRO CB C 32.8 0.3 1 86 9 10 PRO HB2 H 1.82 0.02 2 87 9 10 PRO HB3 H 2.36 0.02 2 88 9 10 PRO CG C 28.0 0.3 1 89 9 10 PRO HG2 H 1.82 0.02 2 90 9 10 PRO HG3 H 2.16 0.02 2 91 9 10 PRO HD2 H 3.63 0.02 2 92 9 10 PRO HD3 H 3.83 0.02 2 93 9 10 PRO C C 174.9 0.3 1 94 10 11 CYS N N 115.0 0.2 1 95 10 11 CYS H H 7.74 0.02 1 96 10 11 CYS CA C 50.9 0.3 1 97 10 11 CYS HA H 5.10 0.02 1 98 10 11 CYS CB C 47.7 0.3 1 99 10 11 CYS HB2 H 2.72 0.02 2 100 10 11 CYS HB3 H 3.39 0.02 2 101 11 12 PRO CD C 50.5 0.3 1 102 11 12 PRO CA C 63.1 0.3 1 103 11 12 PRO HA H 4.39 0.02 1 104 11 12 PRO CB C 31.7 0.3 1 105 11 12 PRO HB2 H 1.54 0.02 2 106 11 12 PRO HB3 H 1.99 0.02 2 107 11 12 PRO CG C 26.3 0.3 1 108 11 12 PRO HG2 H 1.84 0.02 2 109 11 12 PRO HG3 H 2.00 0.02 2 110 11 12 PRO HD2 H 3.40 0.02 2 111 11 12 PRO HD3 H 3.68 0.02 2 112 12 13 TYR CA C 55.9 0.3 1 113 12 13 TYR HA H 4.72 0.02 1 114 12 13 TYR CB C 40.4 0.3 1 115 12 13 TYR HB2 H 2.93 0.02 2 116 12 13 TYR HB3 H 3.41 0.02 2 117 12 13 TYR HD1 H 7.16 0.02 1 118 12 13 TYR HD2 H 7.16 0.02 1 119 12 13 TYR HE1 H 6.82 0.02 1 120 12 13 TYR HE2 H 6.82 0.02 1 121 13 14 ASN N N 119.2 0.2 1 122 13 14 ASN H H 8.77 0.02 1 123 13 14 ASN CA C 56.4 0.3 1 124 13 14 ASN HA H 4.13 0.02 1 125 13 14 ASN CB C 38.5 0.3 1 126 13 14 ASN HB2 H 2.78 0.02 2 127 13 14 ASN HB3 H 2.91 0.02 2 128 13 14 ASN ND2 N 112.3 0.2 1 129 13 14 ASN HD21 H 6.92 0.02 2 130 13 14 ASN HD22 H 7.76 0.02 2 131 14 15 GLU N N 114.5 0.2 1 132 14 15 GLU H H 8.59 0.02 1 133 14 15 GLU CA C 58.0 0.3 1 134 14 15 GLU HA H 4.22 0.02 1 135 14 15 GLU CB C 28.4 0.3 1 136 14 15 GLU HB2 H 2.09 0.02 1 137 14 15 GLU HB3 H 2.09 0.02 1 138 14 15 GLU CG C 33.7 0.3 1 139 14 15 GLU HG3 H 2.49 0.02 2 140 15 16 ASN N N 113.8 0.2 1 141 15 16 ASN H H 7.62 0.02 1 142 15 16 ASN CA C 54.5 0.3 1 143 15 16 ASN HA H 4.70 0.02 1 144 15 16 ASN CB C 39.7 0.3 1 145 15 16 ASN HB2 H 2.49 0.02 2 146 15 16 ASN HB3 H 3.09 0.02 2 147 15 16 ASN ND2 N 109.7 0.2 1 148 15 16 ASN HD21 H 6.91 0.02 2 149 15 16 ASN HD22 H 7.53 0.02 2 150 15 16 ASN C C 174.0 0.3 1 151 16 17 CYS N N 117.9 0.2 1 152 16 17 CYS H H 7.97 0.02 1 153 16 17 CYS CA C 55.7 0.3 1 154 16 17 CYS HA H 5.06 0.02 1 155 16 17 CYS CB C 37.6 0.3 1 156 16 17 CYS HB2 H 2.70 0.02 2 157 16 17 CYS HB3 H 3.42 0.02 2 158 16 17 CYS C C 177.3 0.3 1 159 17 18 CYS N N 128.8 0.2 1 160 17 18 CYS H H 9.74 0.02 1 161 17 18 CYS CA C 57.8 0.3 1 162 17 18 CYS HA H 4.49 0.02 1 163 17 18 CYS CB C 38.7 0.3 1 164 17 18 CYS HB2 H 3.00 0.02 2 165 17 18 CYS HB3 H 3.26 0.02 2 166 17 18 CYS C C 176.6 0.3 1 167 18 19 SER N N 113.8 0.2 1 168 18 19 SER H H 9.20 0.02 1 169 18 19 SER CA C 58.2 0.3 1 170 18 19 SER HA H 4.20 0.02 1 171 18 19 SER CB C 64.7 0.3 1 172 18 19 SER HB2 H 3.78 0.02 2 173 18 19 SER HB3 H 4.00 0.02 2 174 18 19 SER C C 174.9 0.3 1 175 19 20 GLN N N 112.8 0.2 1 176 19 20 GLN H H 7.73 0.02 1 177 19 20 GLN CA C 56.7 0.3 1 178 19 20 GLN HA H 4.04 0.02 1 179 19 20 GLN CB C 26.3 0.3 1 180 19 20 GLN HB2 H 2.33 0.02 1 181 19 20 GLN HB3 H 2.33 0.02 1 182 19 20 GLN CG C 34.2 0.3 1 183 19 20 GLN HG2 H 2.22 0.02 2 184 19 20 GLN HG3 H 2.30 0.02 2 185 19 20 GLN NE2 N 113.7 0.2 1 186 19 20 GLN HE21 H 6.77 0.02 2 187 19 20 GLN HE22 H 7.42 0.02 2 188 20 21 SER N N 111.4 0.2 1 189 20 21 SER H H 7.12 0.02 1 190 20 21 SER CA C 56.9 0.3 1 191 20 21 SER HA H 4.50 0.02 1 192 20 21 SER CB C 63.1 0.3 1 193 20 21 SER HB2 H 3.39 0.02 2 194 20 21 SER HB3 H 3.71 0.02 2 195 20 21 SER C C 172.6 0.3 1 196 21 22 CYS N N 130.2 0.2 1 197 21 22 CYS H H 9.06 0.02 1 198 21 22 CYS CA C 54.9 0.3 1 199 21 22 CYS HA H 4.94 0.02 1 200 21 22 CYS CB C 40.4 0.3 1 201 21 22 CYS HB2 H 2.78 0.02 2 202 21 22 CYS HB3 H 3.06 0.02 2 203 21 22 CYS C C 177.2 0.3 1 204 22 23 THR N N 117.8 0.2 1 205 22 23 THR H H 8.28 0.02 1 206 22 23 THR CA C 59.8 0.3 1 207 22 23 THR HA H 4.71 0.02 1 208 22 23 THR CB C 72.4 0.3 1 209 22 23 THR HB H 4.23 0.02 1 210 22 23 THR HG2 H 1.16 0.02 1 211 22 23 THR CG2 C 21.6 0.3 1 212 22 23 THR C C 174.1 0.3 1 213 23 24 PHE N N 120.9 0.2 1 214 23 24 PHE H H 8.54 0.02 1 215 23 24 PHE CA C 58.8 0.3 1 216 23 24 PHE HA H 4.70 0.02 1 217 23 24 PHE CB C 39.9 0.3 1 218 23 24 PHE HB3 H 2.92 0.02 2 219 23 24 PHE HD1 H 7.16 0.02 1 220 23 24 PHE HD2 H 7.16 0.02 1 221 23 24 PHE HE1 H 7.33 0.02 1 222 23 24 PHE HE2 H 7.33 0.02 1 223 23 24 PHE C C 175.9 0.3 1 224 24 25 LYS N N 123.5 0.2 1 225 24 25 LYS H H 8.73 0.02 1 226 24 25 LYS CA C 54.9 0.3 1 227 24 25 LYS HA H 4.56 0.02 1 228 24 25 LYS CB C 34.5 0.3 1 229 24 25 LYS HB2 H 1.66 0.02 2 230 24 25 LYS HB3 H 1.84 0.02 2 231 24 25 LYS CG C 24.6 0.3 1 232 24 25 LYS HG2 H 1.43 0.02 1 233 24 25 LYS HG3 H 1.43 0.02 1 234 24 25 LYS CD C 28.7 0.3 1 235 24 25 LYS HD2 H 1.66 0.02 1 236 24 25 LYS HD3 H 1.66 0.02 1 237 24 25 LYS CE C 42.0 0.3 1 238 24 25 LYS HE2 H 2.91 0.02 1 239 24 25 LYS HE3 H 2.91 0.02 1 240 24 25 LYS C C 175.5 0.3 1 241 25 26 GLU N N 121.8 0.2 1 242 25 26 GLU H H 8.51 0.02 1 243 25 26 GLU CA C 55.6 0.3 1 244 25 26 GLU HA H 4.58 0.02 1 245 25 26 GLU CB C 29.1 0.3 1 246 25 26 GLU HB2 H 1.95 0.02 2 247 25 26 GLU HB3 H 2.04 0.02 2 248 25 26 GLU CG C 33.0 0.3 1 249 25 26 GLU HG2 H 2.35 0.02 2 250 25 26 GLU HG3 H 2.46 0.02 2 251 25 26 GLU C C 175.9 0.3 1 252 26 27 ASN N N 121.9 0.2 1 253 26 27 ASN H H 8.64 0.02 1 254 26 27 ASN CA C 52.0 0.3 1 255 26 27 ASN HA H 4.85 0.02 1 256 26 27 ASN CB C 39.1 0.3 1 257 26 27 ASN HB2 H 2.87 0.02 2 258 26 27 ASN HB3 H 3.06 0.02 2 259 26 27 ASN ND2 N 112.3 0.2 1 260 26 27 ASN HD21 H 7.06 0.02 2 261 26 27 ASN HD22 H 7.53 0.02 2 262 27 28 GLU N N 119.9 0.2 1 263 27 28 GLU H H 8.67 0.02 1 264 27 28 GLU CA C 57.3 0.3 1 265 27 28 GLU HA H 4.28 0.02 1 266 27 28 GLU CB C 28.3 0.3 1 267 27 28 GLU HB2 H 2.03 0.02 2 268 27 28 GLU HB3 H 2.12 0.02 2 269 27 28 GLU CG C 33.5 0.3 1 270 27 28 GLU HG2 H 2.44 0.02 1 271 27 28 GLU HG3 H 2.44 0.02 1 272 27 28 GLU C C 176.2 0.3 1 273 28 29 ASN N N 117.6 0.2 1 274 28 29 ASN H H 8.27 0.02 1 275 28 29 ASN CA C 53.1 0.3 1 276 28 29 ASN HA H 4.68 0.02 1 277 28 29 ASN CB C 38.4 0.3 1 278 28 29 ASN HB2 H 2.79 0.02 2 279 28 29 ASN HB3 H 2.90 0.02 2 280 28 29 ASN ND2 N 112.2 0.2 1 281 28 29 ASN HD21 H 6.89 0.02 2 282 28 29 ASN HD22 H 7.56 0.02 2 283 28 29 ASN C C 175.4 0.3 1 284 29 30 GLY N N 107.5 0.2 1 285 29 30 GLY H H 8.16 0.02 1 286 29 30 GLY CA C 45.3 0.3 1 287 29 30 GLY HA2 H 3.81 0.02 2 288 29 30 GLY HA3 H 4.16 0.02 2 289 30 31 ASN N N 118.9 0.2 1 290 30 31 ASN H H 7.99 0.02 1 291 30 31 ASN CA C 52.6 0.3 1 292 30 31 ASN HA H 4.89 0.02 1 293 30 31 ASN CB C 39.4 0.3 1 294 30 31 ASN HB2 H 2.70 0.02 2 295 30 31 ASN HB3 H 2.79 0.02 2 296 30 31 ASN ND2 N 112.5 0.2 1 297 30 31 ASN HD21 H 6.96 0.02 2 298 30 31 ASN HD22 H 7.48 0.02 2 299 30 31 ASN C C 174.4 0.3 1 300 31 32 THR N N 117.0 0.2 1 301 31 32 THR H H 8.53 0.02 1 302 31 32 THR CA C 62.2 0.3 1 303 31 32 THR HA H 4.67 0.02 1 304 31 32 THR CB C 69.6 0.3 1 305 31 32 THR HB H 4.07 0.02 1 306 31 32 THR HG2 H 1.09 0.02 1 307 31 32 THR CG2 C 21.9 0.3 1 308 31 32 THR C C 174.2 0.3 1 309 32 33 VAL N N 122.5 0.2 1 310 32 33 VAL H H 8.41 0.02 1 311 32 33 VAL CA C 60.5 0.3 1 312 32 33 VAL HA H 4.49 0.02 1 313 32 33 VAL CB C 35.1 0.3 1 314 32 33 VAL HB H 2.11 0.02 1 315 32 33 VAL HG2 H 0.87 0.02 2 316 32 33 VAL CG2 C 19.8 0.3 1 317 32 33 VAL C C 174.7 0.3 1 318 33 34 LYS N N 126.0 0.2 1 319 33 34 LYS H H 8.39 0.02 1 320 33 34 LYS CA C 56.8 0.3 1 321 33 34 LYS HA H 4.33 0.02 1 322 33 34 LYS CB C 33.3 0.3 1 323 33 34 LYS HB2 H 1.47 0.02 2 324 33 34 LYS HB3 H 1.58 0.02 2 325 33 34 LYS CG C 25.7 0.3 1 326 33 34 LYS HG2 H 0.49 0.02 2 327 33 34 LYS HG3 H 0.71 0.02 2 328 33 34 LYS CD C 29.1 0.3 1 329 33 34 LYS HD3 H 1.24 0.02 2 330 33 34 LYS CE C 41.5 0.3 1 331 33 34 LYS HE2 H 2.45 0.02 2 332 33 34 LYS HE3 H 2.53 0.02 2 333 33 34 LYS C C 176.9 0.3 1 334 34 35 ARG N N 121.8 0.2 1 335 34 35 ARG H H 8.62 0.02 1 336 34 35 ARG CA C 54.4 0.3 1 337 34 35 ARG HA H 4.99 0.02 1 338 34 35 ARG CB C 35.6 0.3 1 339 34 35 ARG HB2 H 1.33 0.02 2 340 34 35 ARG HB3 H 1.39 0.02 2 341 34 35 ARG CG C 27.9 0.3 1 342 34 35 ARG HG2 H 1.19 0.02 2 343 34 35 ARG HG3 H 1.47 0.02 2 344 34 35 ARG CD C 43.0 0.3 1 345 34 35 ARG HD2 H 2.94 0.02 2 346 34 35 ARG HD3 H 3.08 0.02 2 347 34 35 ARG NE N 118.9 0.2 1 348 34 35 ARG HE H 7.36 0.02 1 349 34 35 ARG HH11 H 6.77 0.02 2 350 34 35 ARG C C 176.5 0.3 1 351 35 36 CYS N N 121.0 0.2 1 352 35 36 CYS H H 8.66 0.02 1 353 35 36 CYS CA C 54.1 0.3 1 354 35 36 CYS HA H 5.25 0.02 1 355 35 36 CYS CB C 37.7 0.3 1 356 35 36 CYS HB2 H 2.88 0.02 2 357 35 36 CYS HB3 H 3.52 0.02 2 358 35 36 CYS C C 177.3 0.3 1 359 36 37 ASP N N 131.8 0.2 1 360 36 37 ASP H H 9.24 0.02 1 361 36 37 ASP CA C 56.7 0.3 1 362 36 37 ASP HA H 4.43 0.02 1 363 36 37 ASP CB C 40.7 0.3 1 364 36 37 ASP HB2 H 2.52 0.02 2 365 36 37 ASP HB3 H 2.84 0.02 2 stop_ save_