data_5702 ####################### # 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 5702 _BMRB_flat_file_name bmr5702.str _Entry_type original _Submission_date 2003-02-18 _Accession_date 2003-02-18 _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" 199 "13C chemical shifts" 126 "15N chemical shifts" 40 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2003-03-21 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 4233 'H chemical shift by a different group' stop_ _Original_release_date 2003-02-18 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 Gilles Nicolas . . 3 Maciejewski Mark W. . 4 Zamponi Gerald W. . 5 Menez Andre . . 6 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_label 1 SER 2 PRO 3 THR 4 CYS 5 ILE 6 PRO 7 SER 8 GLY 9 GLN 10 PRO 11 CYS 12 PRO 13 TYR 14 ASN 15 GLU 16 ASN 17 CYS 18 CYS 19 SER 20 GLN 21 SER 22 CYS 23 THR 24 PHE 25 LYS 26 GLU 27 ASN 28 GLU 29 ASN 30 GLY 31 ASN 32 THR 33 VAL 34 LYS 35 ARG 36 CYS 37 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 6192 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_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 n/a 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 . 2 PRO C C 177.3 0.2 1 2 . 3 THR N N 113.7 0.2 1 3 . 3 THR H H 8.28 0.02 1 4 . 3 THR CA C 61.6 0.2 1 5 . 3 THR HA H 4.40 0.02 1 6 . 3 THR CB C 69.6 0.2 1 7 . 3 THR HB H 4.28 0.02 1 8 . 3 THR HG2 H 1.23 0.02 1 9 . 3 THR CG2 C 21.6 0.2 1 10 . 3 THR C C 174.1 0.2 1 11 . 4 CYS N N 119.0 0.2 1 12 . 4 CYS H H 8.03 0.02 1 13 . 4 CYS CA C 54.1 0.2 1 14 . 4 CYS HA H 4.82 0.02 1 15 . 4 CYS CB C 42.5 0.2 1 16 . 4 CYS HB2 H 2.97 0.02 1 17 . 4 CYS HB3 H 3.20 0.02 1 18 . 4 CYS C C 173.4 0.2 1 19 . 5 ILE N N 122.8 0.2 1 20 . 5 ILE H H 9.19 0.02 1 21 . 5 ILE CA C 60.1 0.2 1 22 . 5 ILE HA H 4.41 0.02 1 23 . 5 ILE CB C 40.2 0.2 1 24 . 5 ILE HB H 1.87 0.02 1 25 . 5 ILE HG2 H 1.15 0.02 1 26 . 5 ILE CG2 C 19.1 0.2 1 27 . 5 ILE CG1 C 27.3 0.2 1 28 . 5 ILE HG12 H 1.11 0.02 2 29 . 5 ILE HG13 H 1.78 0.02 2 30 . 5 ILE HD1 H 1.00 0.02 1 31 . 5 ILE CD1 C 14.4 0.2 1 32 . 6 PRO CD C 52.1 0.2 1 33 . 6 PRO CA C 62.4 0.2 1 34 . 6 PRO HA H 4.50 0.02 1 35 . 6 PRO CB C 33.4 0.2 1 36 . 6 PRO HB2 H 2.44 0.02 1 37 . 6 PRO HB3 H 2.44 0.02 1 38 . 6 PRO CG C 27.3 0.2 1 39 . 6 PRO HG2 H 1.97 0.02 2 40 . 6 PRO HG3 H 2.06 0.02 2 41 . 6 PRO HD2 H 3.65 0.02 2 42 . 6 PRO HD3 H 4.24 0.02 2 43 . 6 PRO C C 175.5 0.2 1 44 . 7 SER N N 115.4 0.2 1 45 . 7 SER H H 8.46 0.02 1 46 . 7 SER CA C 61.2 0.2 1 47 . 7 SER HA H 3.83 0.02 1 48 . 7 SER CB C 62.9 0.2 1 49 . 7 SER HB2 H 3.64 0.02 1 50 . 7 SER HB3 H 3.64 0.02 1 51 . 7 SER C C 175.2 0.2 1 52 . 8 GLY N N 112.7 0.2 1 53 . 8 GLY H H 9.53 0.02 1 54 . 8 GLY CA C 44.8 0.2 1 55 . 8 GLY HA2 H 3.66 0.02 1 56 . 8 GLY HA3 H 4.39 0.02 1 57 . 8 GLY C C 174.2 0.2 1 58 . 9 GLN N N 119.4 0.2 1 59 . 9 GLN H H 7.38 0.02 1 60 . 9 GLN CA C 53.3 0.2 1 61 . 9 GLN HA H 4.74 0.02 1 62 . 9 GLN CB C 27.9 0.2 1 63 . 9 GLN HB2 H 1.96 0.02 2 64 . 9 GLN HB3 H 2.11 0.02 2 65 . 9 GLN CG C 33.7 0.2 1 66 . 9 GLN HG2 H 2.19 0.02 2 67 . 9 GLN HG3 H 2.36 0.02 2 68 . 9 GLN NE2 N 113.5 0.2 1 69 . 9 GLN HE21 H 6.94 0.02 2 70 . 9 GLN HE22 H 7.66 0.02 2 71 . 10 PRO CD C 51.0 0.2 1 72 . 10 PRO CA C 63.1 0.2 1 73 . 10 PRO HA H 5.00 0.02 1 74 . 10 PRO CB C 32.8 0.2 1 75 . 10 PRO HB2 H 1.84 0.02 2 76 . 10 PRO HB3 H 2.37 0.02 2 77 . 10 PRO CG C 28.1 0.2 1 78 . 10 PRO HG2 H 1.85 0.02 2 79 . 10 PRO HG3 H 2.17 0.02 2 80 . 10 PRO HD2 H 3.65 0.02 2 81 . 10 PRO HD3 H 3.84 0.02 2 82 . 10 PRO C C 175.0 0.2 1 83 . 11 CYS N N 114.9 0.2 1 84 . 11 CYS H H 7.74 0.02 1 85 . 11 CYS CA C 50.9 0.2 1 86 . 11 CYS HA H 5.07 0.02 1 87 . 11 CYS CB C 48.0 0.2 1 88 . 11 CYS HB2 H 2.66 0.02 1 89 . 11 CYS HB3 H 3.40 0.02 1 90 . 12 PRO CD C 50.5 0.2 1 91 . 12 PRO CA C 63.1 0.2 1 92 . 12 PRO HA H 4.43 0.02 1 93 . 12 PRO CB C 31.7 0.2 1 94 . 12 PRO HB2 H 1.59 0.02 1 95 . 12 PRO HB3 H 1.99 0.02 1 96 . 12 PRO CG C 26.3 0.2 1 97 . 12 PRO HG2 H 1.88 0.02 2 98 . 12 PRO HG3 H 2.03 0.02 2 99 . 12 PRO HD2 H 3.40 0.02 2 100 . 12 PRO HD3 H 3.72 0.02 2 101 . 12 PRO C C 174.0 0.2 1 102 . 13 TYR N N 118.5 0.2 1 103 . 13 TYR H H 8.07 0.02 1 104 . 13 TYR CA C 55.8 0.2 1 105 . 13 TYR HA H 4.77 0.02 1 106 . 13 TYR CB C 41.1 0.2 1 107 . 13 TYR HB2 H 2.95 0.02 2 108 . 13 TYR HB3 H 3.48 0.02 2 109 . 13 TYR HD1 H 7.18 0.02 1 110 . 13 TYR HD2 H 7.18 0.02 1 111 . 13 TYR HE1 H 6.79 0.02 1 112 . 13 TYR HE2 H 6.79 0.02 1 113 . 13 TYR C C 175.8 0.2 1 114 . 14 ASN N N 119.7 0.2 1 115 . 14 ASN H H 8.83 0.02 1 116 . 14 ASN CA C 57.0 0.2 1 117 . 14 ASN HA H 4.02 0.02 1 118 . 14 ASN CB C 38.6 0.2 1 119 . 14 ASN HB2 H 2.76 0.02 2 120 . 14 ASN HB3 H 2.89 0.02 2 121 . 14 ASN ND2 N 112.5 0.2 1 122 . 14 ASN HD21 H 6.91 0.02 2 123 . 14 ASN HD22 H 7.89 0.02 2 124 . 14 ASN C C 177.3 0.2 1 125 . 15 GLU N N 115.3 0.2 1 126 . 15 GLU H H 9.35 0.02 1 127 . 15 GLU CA C 59.3 0.2 1 128 . 15 GLU HA H 4.09 0.02 1 129 . 15 GLU CB C 29.0 0.2 1 130 . 15 GLU HB2 H 2.03 0.02 1 131 . 15 GLU HB3 H 2.03 0.02 1 132 . 15 GLU CG C 37.1 0.2 1 133 . 15 GLU HG2 H 2.31 0.02 2 134 . 15 GLU HG3 H 2.48 0.02 2 135 . 15 GLU C C 176.6 0.2 1 136 . 16 ASN N N 114.2 0.2 1 137 . 16 ASN H H 7.50 0.02 1 138 . 16 ASN CA C 54.5 0.2 1 139 . 16 ASN HA H 4.75 0.02 1 140 . 16 ASN CB C 40.3 0.2 1 141 . 16 ASN HB2 H 2.42 0.02 2 142 . 16 ASN HB3 H 3.08 0.02 2 143 . 16 ASN ND2 N 109.3 0.2 1 144 . 16 ASN HD21 H 6.93 0.02 2 145 . 16 ASN HD22 H 7.51 0.02 2 146 . 16 ASN C C 176.4 0.2 1 147 . 17 CYS N N 118.1 0.2 1 148 . 17 CYS H H 7.91 0.02 1 149 . 17 CYS CA C 55.7 0.2 1 150 . 17 CYS HA H 5.02 0.02 1 151 . 17 CYS CB C 37.9 0.2 1 152 . 17 CYS HB2 H 2.73 0.02 1 153 . 17 CYS HB3 H 3.40 0.02 1 154 . 17 CYS C C 177.2 0.2 1 155 . 18 CYS N N 128.7 0.2 1 156 . 18 CYS H H 9.72 0.02 1 157 . 18 CYS CA C 57.7 0.2 1 158 . 18 CYS HA H 4.48 0.02 1 159 . 18 CYS CB C 38.6 0.2 1 160 . 18 CYS HB2 H 2.98 0.02 1 161 . 18 CYS HB3 H 3.23 0.02 1 162 . 18 CYS C C 176.7 0.2 1 163 . 19 SER N N 113.9 0.2 1 164 . 19 SER H H 9.21 0.02 1 165 . 19 SER CA C 58.2 0.2 1 166 . 19 SER HA H 4.18 0.02 1 167 . 19 SER CB C 64.9 0.2 1 168 . 19 SER HB2 H 3.78 0.02 1 169 . 19 SER HB3 H 3.98 0.02 1 170 . 19 SER C C 175.0 0.2 1 171 . 20 GLN N N 112.8 0.2 1 172 . 20 GLN H H 7.74 0.02 1 173 . 20 GLN CA C 56.8 0.2 1 174 . 20 GLN HA H 4.05 0.02 1 175 . 20 GLN CB C 26.3 0.2 1 176 . 20 GLN HB2 H 2.29 0.02 1 177 . 20 GLN HB3 H 2.29 0.02 1 178 . 20 GLN CG C 34.5 0.2 1 179 . 20 GLN HG2 H 2.18 0.02 2 180 . 20 GLN HG3 H 2.25 0.02 2 181 . 20 GLN NE2 N 114.0 0.2 1 182 . 20 GLN HE21 H 6.75 0.02 2 183 . 20 GLN HE22 H 7.41 0.02 2 184 . 20 GLN C C 174.4 0.2 1 185 . 21 SER N N 111.1 0.2 1 186 . 21 SER H H 7.06 0.02 1 187 . 21 SER CA C 56.9 0.2 1 188 . 21 SER HA H 4.46 0.02 1 189 . 21 SER CB C 62.9 0.2 1 190 . 21 SER HB2 H 3.36 0.02 1 191 . 21 SER HB3 H 3.74 0.02 1 192 . 21 SER C C 170.9 0.2 1 193 . 22 CYS N N 130.9 0.2 1 194 . 22 CYS H H 9.15 0.02 1 195 . 22 CYS CA C 54.9 0.2 1 196 . 22 CYS HA H 4.91 0.02 1 197 . 22 CYS CB C 40.4 0.2 1 198 . 22 CYS HB2 H 3.04 0.02 1 199 . 22 CYS HB3 H 2.73 0.02 1 200 . 22 CYS C C 173.8 0.2 1 201 . 23 THR N N 117.3 0.2 1 202 . 23 THR H H 8.18 0.02 1 203 . 23 THR CA C 59.9 0.2 1 204 . 23 THR HA H 4.72 0.02 1 205 . 23 THR CB C 72.9 0.2 1 206 . 23 THR HB H 4.26 0.02 1 207 . 23 THR HG2 H 1.16 0.02 1 208 . 23 THR CG2 C 21.9 0.2 1 209 . 23 THR C C 174.4 0.2 1 210 . 24 PHE N N 120.9 0.2 1 211 . 24 PHE H H 8.57 0.02 1 212 . 24 PHE CA C 59.0 0.2 1 213 . 24 PHE HA H 4.69 0.02 1 214 . 24 PHE CB C 40.0 0.2 1 215 . 24 PHE HB2 H 2.92 0.02 1 216 . 24 PHE HB3 H 2.92 0.02 1 217 . 24 PHE HD1 H 7.16 0.02 1 218 . 24 PHE HD2 H 7.16 0.02 1 219 . 24 PHE HE1 H 7.33 0.02 1 220 . 24 PHE HE2 H 7.33 0.02 1 221 . 24 PHE C C 176.1 0.2 1 222 . 25 LYS N N 123.3 0.2 1 223 . 25 LYS H H 8.78 0.02 1 224 . 25 LYS CA C 54.6 0.2 1 225 . 25 LYS HA H 4.58 0.02 1 226 . 25 LYS CB C 34.3 0.2 1 227 . 25 LYS HB2 H 1.66 0.02 2 228 . 25 LYS HB3 H 1.87 0.02 2 229 . 25 LYS CG C 24.5 0.2 1 230 . 25 LYS HG2 H 1.43 0.02 1 231 . 25 LYS HG3 H 1.43 0.02 1 232 . 25 LYS CD C 28.6 0.2 1 233 . 25 LYS HD2 H 1.62 0.02 1 234 . 25 LYS HD3 H 1.62 0.02 1 235 . 25 LYS CE C 42.0 0.2 1 236 . 25 LYS HE2 H 2.90 0.02 1 237 . 25 LYS HE3 H 2.90 0.02 1 238 . 25 LYS C C 175.4 0.2 1 239 . 26 GLU N N 122.4 0.2 1 240 . 26 GLU H H 8.52 0.02 1 241 . 26 GLU CA C 56.6 0.2 1 242 . 26 GLU HA H 4.48 0.02 1 243 . 26 GLU CB C 30.7 0.2 1 244 . 26 GLU HB2 H 1.88 0.02 2 245 . 26 GLU HB3 H 1.98 0.02 2 246 . 26 GLU CG C 36.5 0.2 1 247 . 26 GLU HG2 H 2.14 0.02 2 248 . 26 GLU HG3 H 2.24 0.02 2 249 . 26 GLU C C 176.4 0.2 1 250 . 27 ASN N N 121.4 0.2 1 251 . 27 ASN H H 8.65 0.02 1 252 . 27 ASN CA C 52.1 0.2 1 253 . 27 ASN HA H 4.83 0.02 1 254 . 27 ASN CB C 39.5 0.2 1 255 . 27 ASN HB2 H 2.84 0.02 2 256 . 27 ASN HB3 H 3.06 0.02 2 257 . 27 ASN ND2 N 112.6 0.2 1 258 . 27 ASN HD21 H 7.11 0.02 2 259 . 27 ASN HD22 H 7.54 0.02 2 260 . 27 ASN C C 176.1 0.2 1 261 . 28 GLU N N 120.5 0.2 1 262 . 28 GLU H H 8.96 0.02 1 263 . 28 GLU CA C 58.0 0.2 1 264 . 28 GLU HA H 4.22 0.02 1 265 . 28 GLU CB C 29.5 0.2 1 266 . 28 GLU HB2 H 1.96 0.02 2 267 . 28 GLU HB3 H 2.05 0.02 2 268 . 28 GLU CG C 36.1 0.2 1 269 . 28 GLU HG2 H 2.26 0.02 1 270 . 28 GLU HG3 H 2.26 0.02 1 271 . 28 GLU C C 176.5 0.2 1 272 . 29 ASN N N 117.8 0.2 1 273 . 29 ASN H H 8.25 0.02 1 274 . 29 ASN CA C 53.3 0.2 1 275 . 29 ASN HA H 4.76 0.02 1 276 . 29 ASN CB C 38.9 0.2 1 277 . 29 ASN HB2 H 2.73 0.02 2 278 . 29 ASN HB3 H 2.94 0.02 2 279 . 29 ASN ND2 N 112.7 0.2 1 280 . 29 ASN HD21 H 6.91 0.02 2 281 . 29 ASN HD22 H 7.57 0.02 2 282 . 29 ASN C C 175.5 0.2 1 283 . 30 GLY N N 107.5 0.2 1 284 . 30 GLY H H 8.18 0.02 1 285 . 30 GLY CA C 45.3 0.2 1 286 . 30 GLY HA2 H 3.79 0.02 1 287 . 30 GLY HA3 H 4.15 0.02 1 288 . 30 GLY C C 174.0 0.2 1 289 . 31 ASN N N 118.9 0.2 1 290 . 31 ASN H H 7.99 0.02 1 291 . 31 ASN CA C 52.6 0.2 1 292 . 31 ASN HA H 4.90 0.02 1 293 . 31 ASN CB C 39.7 0.2 1 294 . 31 ASN HB2 H 2.68 0.02 2 295 . 31 ASN HB3 H 2.78 0.02 2 296 . 31 ASN ND2 N 112.8 0.2 1 297 . 31 ASN HD21 H 6.96 0.02 2 298 . 31 ASN HD22 H 7.51 0.02 2 299 . 31 ASN C C 174.9 0.2 1 300 . 32 THR N N 117.0 0.2 1 301 . 32 THR H H 8.52 0.02 1 302 . 32 THR CA C 62.1 0.2 1 303 . 32 THR HA H 4.63 0.02 1 304 . 32 THR CB C 69.6 0.2 1 305 . 32 THR HB H 4.04 0.02 1 306 . 32 THR HG2 H 1.07 0.02 1 307 . 32 THR CG2 C 21.9 0.2 1 308 . 32 THR C C 174.3 0.2 1 309 . 33 VAL N N 122.6 0.2 1 310 . 33 VAL H H 8.46 0.02 1 311 . 33 VAL CA C 60.5 0.2 1 312 . 33 VAL HA H 4.49 0.02 1 313 . 33 VAL CB C 35.3 0.2 1 314 . 33 VAL HB H 2.10 0.02 1 315 . 33 VAL HG2 H 0.88 0.02 2 316 . 33 VAL CG2 C 19.8 0.2 1 317 . 33 VAL C C 174.6 0.2 1 318 . 34 LYS N N 126.0 0.2 1 319 . 34 LYS H H 8.38 0.02 1 320 . 34 LYS CA C 56.7 0.2 1 321 . 34 LYS HA H 4.26 0.02 1 322 . 34 LYS CB C 33.4 0.2 1 323 . 34 LYS HB2 H 1.44 0.02 2 324 . 34 LYS HB3 H 1.58 0.02 2 325 . 34 LYS CG C 26.0 0.2 1 326 . 34 LYS HG2 H 0.43 0.02 2 327 . 34 LYS HG3 H 0.59 0.02 2 328 . 34 LYS CD C 29.3 0.2 1 329 . 34 LYS HD2 H 1.13 0.02 2 330 . 34 LYS HD3 H 1.20 0.02 2 331 . 34 LYS CE C 41.7 0.2 1 332 . 34 LYS HE2 H 2.43 0.02 2 333 . 34 LYS HE3 H 2.50 0.02 2 334 . 34 LYS C C 177.0 0.2 1 335 . 35 ARG N N 121.5 0.2 1 336 . 35 ARG H H 8.55 0.02 1 337 . 35 ARG CA C 54.4 0.2 1 338 . 35 ARG HA H 4.99 0.02 1 339 . 35 ARG CB C 35.7 0.2 1 340 . 35 ARG HB2 H 1.30 0.02 2 341 . 35 ARG HB3 H 1.39 0.02 2 342 . 35 ARG CG C 28.1 0.2 1 343 . 35 ARG HG2 H 1.16 0.02 2 344 . 35 ARG HG3 H 1.34 0.02 2 345 . 35 ARG CD C 43.3 0.2 1 346 . 35 ARG HD2 H 2.90 0.02 2 347 . 35 ARG HD3 H 3.08 0.02 2 348 . 35 ARG NE N 119.0 0.2 1 349 . 35 ARG HE H 7.47 0.02 1 350 . 35 ARG C C 176.7 0.2 1 351 . 36 CYS N N 121.0 0.2 1 352 . 36 CYS H H 8.63 0.02 1 353 . 36 CYS CA C 54.3 0.2 1 354 . 36 CYS HA H 5.24 0.02 1 355 . 36 CYS CB C 37.8 0.2 1 356 . 36 CYS HB2 H 2.85 0.02 1 357 . 36 CYS HB3 H 3.52 0.02 1 358 . 36 CYS C C 177.4 0.2 1 359 . 37 ASP N N 132.3 0.2 1 360 . 37 ASP H H 9.29 0.02 1 361 . 37 ASP CA C 57.5 0.2 1 362 . 37 ASP HA H 4.40 0.02 1 363 . 37 ASP CB C 41.7 0.2 1 364 . 37 ASP HB2 H 2.40 0.02 1 365 . 37 ASP HB3 H 2.80 0.02 1 stop_ save_