data_6194 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H, 13C, and 15N Chemical Shift Assignments for Omega-atracotoxin-Hv1a at pH 6.0 ; _BMRB_accession_number 6194 _BMRB_flat_file_name bmr6194.str _Entry_type original _Submission_date 2004-04-26 _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" 206 "13C chemical shifts" 130 "15N chemical shifts" 40 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2004-06-25 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 4233 'H chemical shift by a different group' 5702 'chemical shifts at pH 6.0' 6192 'chemical shifts at pH 3.6' 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 6.0 ; _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 6192 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_3D_HNCACB_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCACB' _Sample_label . save_ save_3D_HNCO_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCO' _Sample_label . save_ save_3D_HNCACO_5 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCACO' _Sample_label . save_ save_3D_HCCH-COSY_6 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HCCH-COSY' _Sample_label . save_ save_3D_(H)CCOHN-TOCSY_7 _Saveframe_category NMR_applied_experiment _Experiment_name '3D (H)CCOHN-TOCSY' _Sample_label . save_ save_3D_1H-15N_TOCSY_8 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-15N TOCSY' _Sample_label . save_ save_3D_HNHA_9 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNHA' _Sample_label . save_ save_3D_HNHB_10 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNHB' _Sample_label . save_ save_2D_1H-1H_NOESY_11 _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' 0.075 0.02 M pH 6.0 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' '3D HNCACB' '3D HNCO' '3D HNCACO' '3D HCCH-COSY' '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 50.8 0.3 1 2 1 2 PRO CA C 63.4 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 2.04 0.02 2 6 1 2 PRO HB3 H 2.38 0.02 2 7 1 2 PRO CG C 27.3 0.3 1 8 1 2 PRO HG2 H 2.08 0.02 1 9 1 2 PRO HG3 H 2.08 0.02 1 10 1 2 PRO HD2 H 3.79 0.02 2 11 1 2 PRO HD3 H 3.87 0.02 2 12 1 2 PRO C C 177.3 0.3 1 13 2 3 THR N N 113.7 0.2 1 14 2 3 THR H H 8.28 0.02 1 15 2 3 THR CA C 61.6 0.3 1 16 2 3 THR HA H 4.40 0.02 1 17 2 3 THR CB C 69.6 0.3 1 18 2 3 THR HB H 4.28 0.02 1 19 2 3 THR HG2 H 1.23 0.02 1 20 2 3 THR CG2 C 21.6 0.3 1 21 2 3 THR C C 174.1 0.3 1 22 3 4 CYS N N 119.0 0.2 1 23 3 4 CYS H H 8.03 0.02 1 24 3 4 CYS CA C 54.1 0.3 1 25 3 4 CYS HA H 4.82 0.02 1 26 3 4 CYS CB C 42.5 0.3 1 27 3 4 CYS HB2 H 2.97 0.02 2 28 3 4 CYS HB3 H 3.20 0.02 2 29 3 4 CYS C C 173.4 0.3 1 30 4 5 ILE N N 122.8 0.2 1 31 4 5 ILE H H 9.19 0.02 1 32 4 5 ILE CA C 60.1 0.3 1 33 4 5 ILE HA H 4.41 0.02 1 34 4 5 ILE CB C 40.2 0.3 1 35 4 5 ILE HB H 1.87 0.02 1 36 4 5 ILE HG2 H 1.15 0.02 1 37 4 5 ILE CG2 C 19.1 0.3 1 38 4 5 ILE CG1 C 27.3 0.3 1 39 4 5 ILE HG12 H 1.11 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 14.4 0.3 1 43 5 6 PRO CD C 52.1 0.3 1 44 5 6 PRO CA C 62.4 0.3 1 45 5 6 PRO HA H 4.50 0.02 1 46 5 6 PRO CB C 33.4 0.3 1 47 5 6 PRO HB2 H 2.44 0.02 1 48 5 6 PRO HB3 H 2.44 0.02 1 49 5 6 PRO CG C 27.3 0.3 1 50 5 6 PRO HG2 H 1.97 0.02 2 51 5 6 PRO HG3 H 2.06 0.02 2 52 5 6 PRO HD2 H 3.65 0.02 2 53 5 6 PRO HD3 H 4.24 0.02 2 54 5 6 PRO C C 175.5 0.3 1 55 6 7 SER N N 115.4 0.2 1 56 6 7 SER H H 8.46 0.02 1 57 6 7 SER CA C 61.2 0.3 1 58 6 7 SER HA H 3.83 0.02 1 59 6 7 SER CB C 62.9 0.3 1 60 6 7 SER HB2 H 3.64 0.02 1 61 6 7 SER HB3 H 3.64 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 9.53 0.02 1 65 7 8 GLY CA C 44.8 0.3 1 66 7 8 GLY HA2 H 3.66 0.02 2 67 7 8 GLY HA3 H 4.39 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 7.38 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 1.96 0.02 2 75 8 9 GLN HB3 H 2.10 0.02 2 76 8 9 GLN CG C 33.7 0.3 1 77 8 9 GLN HG2 H 2.19 0.02 2 78 8 9 GLN HG3 H 2.36 0.02 2 79 8 9 GLN NE2 N 113.5 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 63.1 0.3 1 84 9 10 PRO HA H 5.00 0.02 1 85 9 10 PRO CB C 32.8 0.3 1 86 9 10 PRO HB2 H 1.84 0.02 2 87 9 10 PRO HB3 H 2.37 0.02 2 88 9 10 PRO CG C 28.1 0.3 1 89 9 10 PRO HG2 H 1.85 0.02 2 90 9 10 PRO HG3 H 2.17 0.02 2 91 9 10 PRO HD2 H 3.65 0.02 2 92 9 10 PRO HD3 H 3.84 0.02 2 93 9 10 PRO C C 175.0 0.3 1 94 10 11 CYS N N 114.9 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 5.07 0.02 1 98 10 11 CYS CB C 48.0 0.3 1 99 10 11 CYS HB2 H 2.66 0.02 2 100 10 11 CYS HB3 H 3.40 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 4.43 0.02 1 104 11 12 PRO CB C 31.7 0.3 1 105 11 12 PRO HB2 H 1.59 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 1.88 0.02 2 109 11 12 PRO HG3 H 2.03 0.02 2 110 11 12 PRO HD2 H 3.40 0.02 2 111 11 12 PRO HD3 H 3.72 0.02 2 112 11 12 PRO C C 174.0 0.3 1 113 12 13 TYR N N 118.5 0.2 1 114 12 13 TYR H H 8.07 0.02 1 115 12 13 TYR CA C 55.8 0.3 1 116 12 13 TYR HA H 4.77 0.02 1 117 12 13 TYR CB C 41.1 0.3 1 118 12 13 TYR HB2 H 2.95 0.02 2 119 12 13 TYR HB3 H 3.48 0.02 2 120 12 13 TYR HD1 H 7.18 0.02 1 121 12 13 TYR HD2 H 7.18 0.02 1 122 12 13 TYR HE1 H 6.79 0.02 1 123 12 13 TYR HE2 H 6.79 0.02 1 124 12 13 TYR C C 175.8 0.3 1 125 13 14 ASN N N 119.7 0.2 1 126 13 14 ASN H H 8.83 0.02 1 127 13 14 ASN CA C 57.0 0.3 1 128 13 14 ASN HA H 4.01 0.02 1 129 13 14 ASN CB C 38.6 0.3 1 130 13 14 ASN HB2 H 2.76 0.02 2 131 13 14 ASN HB3 H 2.89 0.02 2 132 13 14 ASN ND2 N 112.5 0.2 1 133 13 14 ASN HD21 H 6.91 0.02 2 134 13 14 ASN HD22 H 7.89 0.02 2 135 13 14 ASN C C 177.3 0.3 1 136 14 15 GLU N N 115.3 0.2 1 137 14 15 GLU H H 9.35 0.02 1 138 14 15 GLU CA C 59.3 0.3 1 139 14 15 GLU HA H 4.09 0.02 1 140 14 15 GLU CB C 29.0 0.3 1 141 14 15 GLU HB2 H 2.03 0.02 1 142 14 15 GLU HB3 H 2.03 0.02 1 143 14 15 GLU CG C 37.1 0.3 1 144 14 15 GLU HG2 H 2.31 0.02 2 145 14 15 GLU HG3 H 2.48 0.02 2 146 14 15 GLU C C 176.6 0.3 1 147 15 16 ASN N N 114.2 0.2 1 148 15 16 ASN H H 7.50 0.02 1 149 15 16 ASN CA C 54.5 0.3 1 150 15 16 ASN HA H 4.75 0.02 1 151 15 16 ASN CB C 40.3 0.3 1 152 15 16 ASN HB2 H 2.42 0.02 2 153 15 16 ASN HB3 H 3.08 0.02 2 154 15 16 ASN ND2 N 109.3 0.2 1 155 15 16 ASN HD21 H 6.93 0.02 2 156 15 16 ASN HD22 H 7.51 0.02 2 157 15 16 ASN C C 176.4 0.3 1 158 16 17 CYS N N 118.1 0.2 1 159 16 17 CYS H H 7.91 0.02 1 160 16 17 CYS CA C 55.6 0.3 1 161 16 17 CYS HA H 5.02 0.02 1 162 16 17 CYS CB C 37.9 0.3 1 163 16 17 CYS HB2 H 2.73 0.02 2 164 16 17 CYS HB3 H 3.40 0.02 2 165 16 17 CYS C C 177.2 0.3 1 166 17 18 CYS N N 128.7 0.2 1 167 17 18 CYS H H 9.72 0.02 1 168 17 18 CYS CA C 57.7 0.3 1 169 17 18 CYS HA H 4.48 0.02 1 170 17 18 CYS CB C 38.6 0.3 1 171 17 18 CYS HB2 H 2.98 0.02 2 172 17 18 CYS HB3 H 3.23 0.02 2 173 17 18 CYS C C 176.7 0.3 1 174 18 19 SER N N 113.9 0.2 1 175 18 19 SER H H 9.21 0.02 1 176 18 19 SER CA C 58.2 0.3 1 177 18 19 SER HA H 4.18 0.02 1 178 18 19 SER CB C 64.9 0.3 1 179 18 19 SER HB2 H 3.78 0.02 2 180 18 19 SER HB3 H 3.98 0.02 2 181 18 19 SER C C 175.0 0.3 1 182 19 20 GLN N N 112.8 0.2 1 183 19 20 GLN H H 7.74 0.02 1 184 19 20 GLN CA C 56.8 0.3 1 185 19 20 GLN HA H 4.05 0.02 1 186 19 20 GLN CB C 26.3 0.3 1 187 19 20 GLN HB2 H 2.29 0.02 1 188 19 20 GLN HB3 H 2.29 0.02 1 189 19 20 GLN CG C 34.5 0.3 1 190 19 20 GLN HG2 H 2.18 0.02 2 191 19 20 GLN HG3 H 2.25 0.02 2 192 19 20 GLN NE2 N 114.0 0.2 1 193 19 20 GLN HE21 H 6.75 0.02 2 194 19 20 GLN HE22 H 7.41 0.02 2 195 19 20 GLN C C 174.4 0.3 1 196 20 21 SER N N 111.1 0.2 1 197 20 21 SER H H 7.06 0.02 1 198 20 21 SER CA C 56.9 0.3 1 199 20 21 SER HA H 4.46 0.02 1 200 20 21 SER CB C 62.9 0.3 1 201 20 21 SER HB2 H 3.36 0.02 2 202 20 21 SER HB3 H 3.73 0.02 2 203 20 21 SER C C 170.9 0.3 1 204 21 22 CYS N N 130.9 0.2 1 205 21 22 CYS H H 9.15 0.02 1 206 21 22 CYS CA C 54.9 0.3 1 207 21 22 CYS HA H 4.91 0.02 1 208 21 22 CYS CB C 40.4 0.3 1 209 21 22 CYS HB2 H 2.73 0.02 2 210 21 22 CYS HB3 H 3.04 0.02 2 211 21 22 CYS C C 173.8 0.3 1 212 22 23 THR N N 117.3 0.2 1 213 22 23 THR H H 8.18 0.02 1 214 22 23 THR CA C 59.9 0.3 1 215 22 23 THR HA H 4.71 0.02 1 216 22 23 THR CB C 72.9 0.3 1 217 22 23 THR HB H 4.25 0.02 1 218 22 23 THR HG2 H 1.16 0.02 1 219 22 23 THR CG2 C 21.9 0.3 1 220 22 23 THR C C 174.4 0.3 1 221 23 24 PHE N N 120.9 0.2 1 222 23 24 PHE H H 8.56 0.02 1 223 23 24 PHE CA C 59.0 0.3 1 224 23 24 PHE HA H 4.69 0.02 1 225 23 24 PHE CB C 40.0 0.3 1 226 23 24 PHE HB2 H 2.91 0.02 1 227 23 24 PHE HB3 H 2.91 0.02 1 228 23 24 PHE HD1 H 7.16 0.02 1 229 23 24 PHE HD2 H 7.16 0.02 1 230 23 24 PHE HE1 H 7.33 0.02 1 231 23 24 PHE HE2 H 7.33 0.02 1 232 23 24 PHE C C 176.1 0.3 1 233 24 25 LYS N N 123.3 0.2 1 234 24 25 LYS H H 8.78 0.02 1 235 24 25 LYS CA C 54.6 0.3 1 236 24 25 LYS HA H 4.58 0.02 1 237 24 25 LYS CB C 34.3 0.3 1 238 24 25 LYS HB2 H 1.66 0.02 2 239 24 25 LYS HB3 H 1.87 0.02 2 240 24 25 LYS CG C 24.5 0.3 1 241 24 25 LYS HG2 H 1.43 0.02 1 242 24 25 LYS HG3 H 1.43 0.02 1 243 24 25 LYS CD C 28.6 0.3 1 244 24 25 LYS HD2 H 1.62 0.02 1 245 24 25 LYS HD3 H 1.62 0.02 1 246 24 25 LYS CE C 42.0 0.3 1 247 24 25 LYS HE2 H 2.90 0.02 1 248 24 25 LYS HE3 H 2.90 0.02 1 249 24 25 LYS C C 175.4 0.3 1 250 25 26 GLU N N 122.4 0.2 1 251 25 26 GLU H H 8.52 0.02 1 252 25 26 GLU CA C 56.6 0.3 1 253 25 26 GLU HA H 4.48 0.02 1 254 25 26 GLU CB C 30.7 0.3 1 255 25 26 GLU HB2 H 1.88 0.02 2 256 25 26 GLU HB3 H 1.98 0.02 2 257 25 26 GLU CG C 36.5 0.3 1 258 25 26 GLU HG2 H 2.14 0.02 2 259 25 26 GLU HG3 H 2.24 0.02 2 260 25 26 GLU C C 176.4 0.3 1 261 26 27 ASN N N 121.4 0.2 1 262 26 27 ASN H H 8.65 0.02 1 263 26 27 ASN CA C 52.1 0.3 1 264 26 27 ASN HA H 4.83 0.02 1 265 26 27 ASN CB C 39.5 0.3 1 266 26 27 ASN HB2 H 2.84 0.02 2 267 26 27 ASN HB3 H 3.06 0.02 2 268 26 27 ASN ND2 N 112.6 0.2 1 269 26 27 ASN HD21 H 7.11 0.02 2 270 26 27 ASN HD22 H 7.54 0.02 2 271 26 27 ASN C C 176.1 0.3 1 272 27 28 GLU N N 120.5 0.2 1 273 27 28 GLU H H 8.96 0.02 1 274 27 28 GLU CA C 58.0 0.3 1 275 27 28 GLU HA H 4.22 0.02 1 276 27 28 GLU CB C 29.5 0.3 1 277 27 28 GLU HB2 H 1.96 0.02 2 278 27 28 GLU HB3 H 2.04 0.02 2 279 27 28 GLU CG C 36.1 0.3 1 280 27 28 GLU HG2 H 2.26 0.02 1 281 27 28 GLU HG3 H 2.26 0.02 1 282 27 28 GLU C C 176.5 0.3 1 283 28 29 ASN N N 117.8 0.2 1 284 28 29 ASN H H 8.25 0.02 1 285 28 29 ASN CA C 53.3 0.3 1 286 28 29 ASN HA H 4.76 0.02 1 287 28 29 ASN CB C 38.9 0.3 1 288 28 29 ASN HB2 H 2.73 0.02 2 289 28 29 ASN HB3 H 2.94 0.02 2 290 28 29 ASN ND2 N 112.7 0.2 1 291 28 29 ASN HD21 H 6.91 0.02 2 292 28 29 ASN HD22 H 7.57 0.02 2 293 28 29 ASN C C 175.5 0.3 1 294 29 30 GLY N N 107.5 0.2 1 295 29 30 GLY H H 8.18 0.02 1 296 29 30 GLY CA C 45.3 0.3 1 297 29 30 GLY HA2 H 3.79 0.02 2 298 29 30 GLY HA3 H 4.15 0.02 2 299 29 30 GLY C C 174.0 0.3 1 300 30 31 ASN N N 118.9 0.2 1 301 30 31 ASN H H 7.99 0.02 1 302 30 31 ASN CA C 52.6 0.3 1 303 30 31 ASN HA H 4.90 0.02 1 304 30 31 ASN CB C 39.7 0.3 1 305 30 31 ASN HB2 H 2.68 0.02 2 306 30 31 ASN HB3 H 2.78 0.02 2 307 30 31 ASN ND2 N 112.8 0.2 1 308 30 31 ASN HD21 H 6.96 0.02 2 309 30 31 ASN HD22 H 7.51 0.02 2 310 30 31 ASN C C 174.9 0.3 1 311 31 32 THR N N 117.0 0.2 1 312 31 32 THR H H 8.52 0.02 1 313 31 32 THR CA C 62.1 0.3 1 314 31 32 THR HA H 4.63 0.02 1 315 31 32 THR CB C 69.6 0.3 1 316 31 32 THR HB H 4.04 0.02 1 317 31 32 THR HG2 H 1.07 0.02 1 318 31 32 THR CG2 C 21.9 0.3 1 319 31 32 THR C C 174.3 0.3 1 320 32 33 VAL N N 122.6 0.2 1 321 32 33 VAL H H 8.46 0.02 1 322 32 33 VAL CA C 60.5 0.3 1 323 32 33 VAL HA H 4.49 0.02 1 324 32 33 VAL CB C 35.3 0.3 1 325 32 33 VAL HB H 2.10 0.02 1 326 32 33 VAL HG2 H 0.88 0.02 2 327 32 33 VAL CG2 C 19.8 0.3 1 328 32 33 VAL C C 174.6 0.3 1 329 33 34 LYS N N 126.0 0.2 1 330 33 34 LYS H H 8.38 0.02 1 331 33 34 LYS CA C 56.7 0.3 1 332 33 34 LYS HA H 4.26 0.02 1 333 33 34 LYS CB C 33.4 0.3 1 334 33 34 LYS HB2 H 1.44 0.02 2 335 33 34 LYS HB3 H 1.58 0.02 2 336 33 34 LYS CG C 26.0 0.3 1 337 33 34 LYS HG2 H 0.43 0.02 2 338 33 34 LYS HG3 H 0.59 0.02 2 339 33 34 LYS CD C 29.3 0.3 1 340 33 34 LYS HD2 H 1.13 0.02 2 341 33 34 LYS HD3 H 1.20 0.02 2 342 33 34 LYS CE C 41.7 0.3 1 343 33 34 LYS HE2 H 2.43 0.02 2 344 33 34 LYS HE3 H 2.50 0.02 2 345 33 34 LYS C C 177.0 0.3 1 346 34 35 ARG N N 121.5 0.2 1 347 34 35 ARG H H 8.55 0.02 1 348 34 35 ARG CA C 54.4 0.3 1 349 34 35 ARG HA H 4.99 0.02 1 350 34 35 ARG CB C 35.7 0.3 1 351 34 35 ARG HB2 H 1.30 0.02 2 352 34 35 ARG HB3 H 1.39 0.02 2 353 34 35 ARG CG C 28.1 0.3 1 354 34 35 ARG HG2 H 1.16 0.02 2 355 34 35 ARG HG3 H 1.34 0.02 2 356 34 35 ARG CD C 43.3 0.3 1 357 34 35 ARG HD2 H 2.90 0.02 2 358 34 35 ARG HD3 H 3.08 0.02 2 359 34 35 ARG NE N 119.0 0.2 1 360 34 35 ARG HE H 7.47 0.02 1 361 34 35 ARG C C 176.7 0.3 1 362 35 36 CYS N N 121.0 0.2 1 363 35 36 CYS H H 8.63 0.02 1 364 35 36 CYS CA C 54.3 0.3 1 365 35 36 CYS HA H 5.24 0.02 1 366 35 36 CYS CB C 37.8 0.3 1 367 35 36 CYS HB2 H 2.85 0.02 2 368 35 36 CYS HB3 H 3.52 0.02 2 369 35 36 CYS C C 177.4 0.3 1 370 36 37 ASP N N 132.3 0.2 1 371 36 37 ASP H H 9.29 0.02 1 372 36 37 ASP CA C 57.5 0.3 1 373 36 37 ASP HA H 4.39 0.02 1 374 36 37 ASP CB C 41.7 0.3 1 375 36 37 ASP HB2 H 2.40 0.02 2 376 36 37 ASP HB3 H 2.80 0.02 2 stop_ save_