data_5240 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 15N assignment of Alicyclobacillus acidocaldarius thermostable thioredoxin ; _BMRB_accession_number 5240 _BMRB_flat_file_name bmr5240.str _Entry_type original _Submission_date 2001-12-20 _Accession_date 2001-12-20 _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 Leone Marilisa . . 2 'Di Lello' Paola . . 3 Pedone Emilia Maria . 4 Bartolucci Simonetta . . 5 Rossi Mose' . . 6 'Di Blasio' Benedetto . . 7 Pedone Carlo . . 8 Saviano Michele . . 9 Isernia Carla . . 10 Fattorusso Roberto . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "15N chemical shifts" 111 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2004-09-10 original author . stop_ loop_ _Related_BMRB_accession_number _Relationship 5241 'Alicyclobacillus acidocaldarius thioredoxin (K18G/R82E mutant)' stop_ _Original_release_date 2004-09-10 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Solution structure and backbone dynamics of the K18G/R82E Alicyclobacillus acidocaldarius thioredoxin mutant: a molecular analysis of its reduced thermal stability. ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 15147188 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Leone Marilisa . . 2 'Di Lello' Paola . . 3 Ohlenschlager O. . . 4 Pedone Emilia Maria . 5 Bartolucci Simonetta . . 6 Rossi Mose' . . 7 'Di Blasio' Benedetto . . 8 Pedone Carlo . . 9 Saviano Michele . . 10 Isernia Carla . . 11 Fattorusso Roberto . . stop_ _Journal_abbreviation Biochemistry _Journal_volume 43 _Journal_issue 20 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 6043 _Page_last 6058 _Year 2004 _Details . save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Pedone E, Saviano M, Rossi M, Bartolucci S. A single point mutation (Glu85Arg) increases the stability of the thioredoxin from Escherichia coli. Protein Eng. 2001 Apr;14(4):255-60. ; _Citation_title 'A single point mutation (Glu85Arg) increases the stability of the thioredoxin from Escherichia coli.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11391017 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Pedone E . . 2 Saviano M . . 3 Rossi M . . 4 Bartolucci S . . stop_ _Journal_abbreviation 'Protein Eng.' _Journal_name_full 'Protein engineering' _Journal_volume 14 _Journal_issue 4 _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 255 _Page_last 260 _Year 2001 _Details ; Glu85 in the Escherichia coli thioredoxin, which is localized in the loop between beta4 and beta5, was substituted with the Arg present in the corresponding position in Bacillus acidocaldarius thioredoxin. This suggested that it could play an important role in the structure and thermostability of this protein owing to its involvement in numerous interactions. The effects of the mutation on the biophysical properties were analysed by circular dichroism, spectrofluorimetry and limited proteolysis, supported by molecular dynamics data. As modelling predicted, an increase in stability for E85R due to additional H-bonds between the beta5 and alpha4 regions was observed. ; save_ save_ref_2 _Saveframe_category citation _Citation_full ; Nicastro G, De Chiara C, Pedone E, Tato M, Rossi M, Bartolucci S. NMR solution structure of a novel thioredoxin from Bacillus acidocaldarius possible determinants of protein stability. Eur J Biochem. 2000 Jan;267(2):403-13. ; _Citation_title 'NMR solution structure of a novel thioredoxin from Bacillus acidocaldarius possible determinants of protein stability.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 10632710 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Nicastro G . . 2 'De Chiara' C . . 3 Pedone E . . 4 Tato M . . 5 Rossi M . . 6 Bartolucci S . . stop_ _Journal_abbreviation 'Eur. J. Biochem.' _Journal_name_full 'European journal of biochemistry / FEBS' _Journal_volume 267 _Journal_issue 2 _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 403 _Page_last 413 _Year 2000 _Details ; The thioredoxin (Trx) from Bacillus acidocaldarius (BacTrx), an eubacterium growing optimally at 333 K, is the first Trx described to date from a moderate thermophilic source. To understand the molecular basis of its thermostability, the three-dimensional structure in the oxidized form was determined by NMR methods. A total of 2276 1H-NMR derived distance constraints along with 23 hydrogen-bonds, 72 phi and 27 chi1 torsion angle restraints, were used in a protocol employing simulated annealing followed by restrained molecular dynamics and restrained energy minimization. BacTrx consists of a well-defined core region of five strands of beta-sheet, surrounded by four exposed alpha-helices, features shared by other members of the thioredoxin family. The BacTrx 3D structure was compared with the Escherichia coli Trx (EcTrx) determined by X-ray crystallographic diffraction, and a number of structural differences were observed that may contribute to its thermostabilty. The results of structural analysis indicated that protein stability is due to cumulative effects, the main factor being an increased number of ionic interactions cross-linking different secondary structural elements and clamping the C-terminal alpha-helix to the core of the protein. ; save_ save_ref_3 _Saveframe_category citation _Citation_full ; Bartolucci S, Guagliardi A, Pedone E, De Pascale D, Cannio R, Camardella L, Rossi M, Nicastro G, de Chiara C, Facci P, Mascetti G, Nicolini C. Thioredoxin from Bacillus acidocaldarius: characterization, high-level expression in Escherichia coli and molecular modelling. Biochem J. 1997 Nov 15;328 ( Pt 1):277-85. ; _Citation_title 'Thioredoxin from Bacillus acidocaldarius: characterization, high-level expression in Escherichia coli and molecular modelling.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 9359865 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Bartolucci S . . 2 Guagliardi A . . 3 Pedone E . . 4 'De Pascale' D . . 5 Cannio R . . 6 Camardella L . . 7 Rossi M . . 8 Nicastro G . . 9 'de Chiara' C . . 10 Facci P . . 11 Mascetti G . . 12 Nicolini C . . stop_ _Journal_abbreviation 'Biochem. J.' _Journal_name_full 'The Biochemical journal' _Journal_volume '328 ( Pt 1)' _Journal_issue . _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 277 _Page_last 285 _Year 1997 _Details ; The thioredoxin (Trx) from Bacillus acidocaldarius (BacTrx) was purified to homogeneity by anion-exchange chromatography and gel-filtration chromatography, based on its ability to catalyse the dithiothreitol-dependent reduction of bovine insulin disulphides. The protein has a molecular mass of 11577 Da, determined by electrospray mass spectrometry, a pI of 4.2, and its primary structure was obtained by automated Edman degradation after cleavage with trypsin and cyanogen bromide. The sequences of known bacterial Trxs were aligned at the active site: BacTrx has an identity ranging from 45 to 53% with all sequences except that of the unusual Anabaena strain 7120 Trx (37% identity). The gene coding for BacTrx was isolated by a strategy based on PCR gene amplification and cloned in a plasmid downstream of a lac-derived promoter sequence; the recombinant clone was used as the expression vector for Escherichia coli. The expression was optimized by varying both the time of cell growth and the time of exposure to the inducer isopropyl beta-d-thiogalactoside; expressed BacTrx represents approx. 5% of the total cytosolic protein. CD spectra and differential scanning calorimetry measurements demonstrated that BacTrx is endowed with a higher conformational heat stability than the Trx from E. coli. Nanogravimetry experiments showed a lower content of bound water in BacTrx than in E. coli Trx, and a transition temperature approx. 10 degrees C higher for BacTrx. The three-dimensional model of the oxidized form of BacTrx was constructed by a comparative molecular modelling technique, using E. coli Trx and Anabaena strain 7120 Trx as reference proteins. Increased networks of ion-pairs and shorter loops emerged as major features of the BacTrx structure compared with those of the template proteins. The findings are discussed in the light of the current knowledge about molecular determinants of protein stability. ; save_ ################################## # Molecular system description # ################################## save_system_Trx _Saveframe_category molecular_system _Mol_system_name Thioredoxin _Abbreviation_common Trx _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'BacTrx wt' $Trx_wt stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all disulfide bound' loop_ _Biological_function 'disulfide oxidoreductase' stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_Trx_wt _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'Alicyclobacillus acidocaldarius thioredoxin' _Abbreviation_common 'BacTrx wt' _Molecular_mass 11573.2 _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 105 _Mol_residue_sequence ; ATMTLTDANFQQAIQGDKPV LVDFWAAWCGPCRMMAPVLE EFAEAHADKVTVAKLNVDEN PETTSQFGIMSIPTLILFKG GRPVKQLIGYQPKEQLEAQL ADVLQ ; loop_ _Residue_seq_code _Residue_label 1 ALA 2 THR 3 MET 4 THR 5 LEU 6 THR 7 ASP 8 ALA 9 ASN 10 PHE 11 GLN 12 GLN 13 ALA 14 ILE 15 GLN 16 GLY 17 ASP 18 LYS 19 PRO 20 VAL 21 LEU 22 VAL 23 ASP 24 PHE 25 TRP 26 ALA 27 ALA 28 TRP 29 CYS 30 GLY 31 PRO 32 CYS 33 ARG 34 MET 35 MET 36 ALA 37 PRO 38 VAL 39 LEU 40 GLU 41 GLU 42 PHE 43 ALA 44 GLU 45 ALA 46 HIS 47 ALA 48 ASP 49 LYS 50 VAL 51 THR 52 VAL 53 ALA 54 LYS 55 LEU 56 ASN 57 VAL 58 ASP 59 GLU 60 ASN 61 PRO 62 GLU 63 THR 64 THR 65 SER 66 GLN 67 PHE 68 GLY 69 ILE 70 MET 71 SER 72 ILE 73 PRO 74 THR 75 LEU 76 ILE 77 LEU 78 PHE 79 LYS 80 GLY 81 GLY 82 ARG 83 PRO 84 VAL 85 LYS 86 GLN 87 LEU 88 ILE 89 GLY 90 TYR 91 GLN 92 PRO 93 LYS 94 GLU 95 GLN 96 LEU 97 GLU 98 ALA 99 GLN 100 LEU 101 ALA 102 ASP 103 VAL 104 LEU 105 GLN stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-01-28 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 4446 Bactrx 100.00 105 100.00 100.00 2.01e-70 BMRB 5241 Trx_mut 100.00 105 98.10 98.10 1.78e-68 PDB 1NSW "The Crystal Structure Of The K18g Mutant Of The Thioredoxin From Alicyclobacillus Acidocaldarius" 100.00 105 99.05 99.05 2.13e-69 PDB 1NW2 "The Crystal Structure Of The Mutant R82e Of Thioredoxin From Alicyclobacillus Acidocaldarius" 100.00 105 99.05 99.05 1.83e-69 PDB 1QUW "Solution Structure Of The Thioredoxin From Bacillus Acidocaldarius" 100.00 105 100.00 100.00 2.01e-70 PDB 1RQM "Solution Structure Of The K18gR82E ALICYCLOBACILLUS Acidocaldarius Thioredoxin Mutant" 100.00 105 98.10 98.10 1.78e-68 GB ACV57898 "thioredoxin [Alicyclobacillus acidocaldarius subsp. acidocaldarius DSM 446]" 100.00 106 100.00 100.00 1.58e-70 GB AEJ42820 "thioredoxin [Alicyclobacillus acidocaldarius subsp. acidocaldarius Tc-4-1]" 98.10 103 99.03 100.00 1.42e-68 GB EED08563 "thioredoxin [Alicyclobacillus acidocaldarius LAA1]" 100.00 106 98.10 98.10 1.24e-68 REF WP_008336730 "thioredoxin [Alicyclobacillus acidocaldarius]" 100.00 106 98.10 98.10 1.24e-68 REF WP_012810252 "thioredoxin [Alicyclobacillus acidocaldarius]" 100.00 106 100.00 100.00 1.58e-70 REF WP_014463721 "thioredoxin [Alicyclobacillus acidocaldarius]" 98.10 103 99.03 100.00 1.42e-68 REF YP_003184287 "thioredoxin [Alicyclobacillus acidocaldarius subsp. acidocaldarius DSM 446]" 100.00 106 100.00 100.00 1.58e-70 REF YP_005517339 "thioredoxin [Alicyclobacillus acidocaldarius subsp. acidocaldarius Tc-4-1]" 98.10 103 99.03 100.00 1.42e-68 SP P80579 "RecName: Full=Thioredoxin; Short=Trx [Alicyclobacillus acidocaldarius subsp. acidocaldarius]" 100.00 105 100.00 100.00 2.01e-70 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $Trx_wt 'Alicyclobacillus acidocaldarius (Bacillus acidocaldarius)' 1388 Eubacteria . Alicyclobacillus acidocaldarius stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Vector_type _Vector_name $Trx_wt 'recombinant technology' 'E. coli' Escherichia coli Rb791 plasmid pTrc99A stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $Trx_wt 1.0 mM [U-15N] stop_ save_ ############################ # Computer software used # ############################ save_Vnmr _Saveframe_category software _Name VNMR _Version 6.1B loop_ _Task 'spectra processing' stop_ _Details . save_ save_PROSA _Saveframe_category software _Name PROSA _Version . loop_ _Task 'spectra processing' stop_ _Details . save_ save_xeasy _Saveframe_category software _Name XEASY _Version 1.3.13 loop_ _Task 'data analysis' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model INOVA _Field_strength 750 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model UNITY_INOVA _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-1H_DQCOSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H DQCOSY' _Sample_label . save_ save_2D_1H-1H_NOESY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label . save_ save_2D_1H-1H_TOCSY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label . save_ save_2D_1H-15N_HSQC_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label . save_ save_3D_1H-1H-15N_NOESY_5 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-1H-15N NOESY' _Sample_label . save_ save_3D_1H-1H-15N_TOCSY_6 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-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_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H DQCOSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_5 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-1H-15N NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_6 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-1H-15N TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_7 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNHA' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_8 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNHB' _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_condition_BacTrx _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 6.0 0.1 na temperature 298.0 0.1 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 water H 1 protons ppm 4.75 internal direct spherical internal parallel 1.0 'ammonium chloride' N 15 nitrogen ppm 119.33647 external direct spherical external parallel 1.0 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_shifts_BacTrx_wt _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-1H DQCOSY' '2D 1H-1H NOESY' '2D 1H-1H TOCSY' '2D 1H-15N HSQC' '3D 1H-1H-15N NOESY' '3D 1H-1H-15N TOCSY' '3D HNHA' '3D HNHB' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $condition_BacTrx _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'BacTrx wt' _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 THR N N 121.9 0.1 1 2 . 3 MET N N 124.2 0.1 1 3 . 4 THR N N 123.8 0.1 1 4 . 5 LEU N N 129.3 0.1 1 5 . 6 THR N N 111.8 0.1 1 6 . 7 ASP N N 121.1 0.1 1 7 . 8 ALA N N 121.0 0.1 1 8 . 9 ASN N N 116.5 0.1 1 9 . 9 ASN ND2 N 107.2 0.1 1 10 . 10 PHE N N 124.1 0.1 1 11 . 11 GLN N N 116.1 0.1 1 12 . 11 GLN NE2 N 113.0 0.1 1 13 . 12 GLN N N 117.5 0.1 1 14 . 12 GLN NE2 N 113.0 0.1 1 15 . 13 ALA N N 121.8 0.1 1 16 . 14 ILE N N 109.4 0.1 1 17 . 15 GLN N N 120.4 0.1 1 18 . 15 GLN NE2 N 113.5 0.1 1 19 . 16 GLY N N 109.2 0.1 1 20 . 17 ASP N N 119.0 0.1 1 21 . 18 LYS N N 122.7 0.1 1 22 . 20 VAL N N 126.1 0.1 1 23 . 21 LEU N N 132.6 0.1 1 24 . 22 VAL N N 128.2 0.1 1 25 . 23 ASP N N 126.7 0.1 1 26 . 24 PHE N N 130.6 0.1 1 27 . 25 TRP N N 122.0 0.1 1 28 . 25 TRP NE1 N 132.5 0.1 1 29 . 26 ALA N N 117.7 0.1 1 30 . 27 ALA N N 125.1 0.1 1 31 . 28 TRP N N 113.4 0.1 1 32 . 28 TRP NE1 N 138.3 0.1 1 33 . 29 CYS N N 122.7 0.1 1 34 . 30 GLY N N 123.6 0.1 1 35 . 32 CYS N N 112.7 0.1 1 36 . 33 ARG N N 123.5 0.1 1 37 . 34 MET N N 119.0 0.1 1 38 . 35 MET N N 117.2 0.1 1 39 . 36 ALA N N 124.3 0.1 1 40 . 38 VAL N N 121.3 0.1 1 41 . 39 LEU N N 121.3 0.1 1 42 . 40 GLU N N 119.0 0.1 1 43 . 41 GLU N N 121.0 0.1 1 44 . 42 PHE N N 122.9 0.1 1 45 . 43 ALA N N 121.9 0.1 1 46 . 44 GLU N N 117.7 0.1 1 47 . 45 ALA N N 121.3 0.1 1 48 . 46 HIS N N 113.5 0.1 1 49 . 47 ALA N N 125.4 0.1 1 50 . 48 ASP N N 115.0 0.1 1 51 . 49 LYS N N 119.3 0.1 1 52 . 50 VAL N N 116.7 0.1 1 53 . 51 THR N N 124.2 0.1 1 54 . 52 VAL N N 130.0 0.1 1 55 . 53 ALA N N 131.2 0.1 1 56 . 54 LYS N N 119.0 0.1 1 57 . 55 LEU N N 125.4 0.1 1 58 . 56 ASN N N 127.4 0.1 1 59 . 56 ASN ND2 N 111.3 0.1 1 60 . 57 VAL N N 124.6 0.1 1 61 . 58 ASP N N 122.7 0.1 1 62 . 59 GLU N N 117.0 0.1 1 63 . 60 ASN N N 117.2 0.1 1 64 . 60 ASN ND2 N 123.1 0.1 1 65 . 62 GLU N N 123.5 0.1 1 66 . 63 THR N N 122.3 0.1 1 67 . 64 THR N N 117.7 0.1 1 68 . 65 SER N N 115.5 0.1 1 69 . 66 GLN N N 124.9 0.1 1 70 . 66 GLN NE2 N 113.0 0.1 1 71 . 67 PHE N N 113.8 0.1 1 72 . 68 GLY N N 110.9 0.1 1 73 . 69 ILE N N 119.0 0.1 1 74 . 70 MET N N 128.9 0.1 1 75 . 71 SER N N 116.6 0.1 1 76 . 72 ILE N N 118.4 0.1 1 77 . 74 THR N N 121.1 0.1 1 78 . 75 LEU N N 128.7 0.1 1 79 . 76 ILE N N 121.7 0.1 1 80 . 77 LEU N N 130.0 0.1 1 81 . 78 PHE N N 129.3 0.1 1 82 . 79 LYS N N 120.3 0.1 1 83 . 80 GLY N N 120.1 0.1 1 84 . 81 GLY N N 106.2 0.1 1 85 . 82 ARG N N 119.7 0.1 1 86 . 84 VAL N N 122.4 0.1 1 87 . 85 LYS N N 120.0 0.1 1 88 . 86 GLN N N 124.9 0.1 1 89 . 86 GLN NE2 N 110.5 0.1 1 90 . 87 LEU N N 127.1 0.1 1 91 . 88 ILE N N 125.3 0.1 1 92 . 89 GLY N N 114.8 0.1 1 93 . 90 TYR N N 119.7 0.1 1 94 . 91 GLN N N 129.2 0.1 1 95 . 91 GLN NE2 N 111.5 0.1 1 96 . 93 LYS N N 124.4 0.1 1 97 . 94 GLU N N 116.6 0.1 1 98 . 95 GLN N N 120.8 0.1 1 99 . 95 GLN NE2 N 113.5 0.1 1 100 . 96 LEU N N 123.2 0.1 1 101 . 97 GLU N N 115.9 0.1 1 102 . 98 ALA N N 120.9 0.1 1 103 . 99 GLN N N 116.4 0.1 1 104 . 99 GLN NE2 N 112.0 0.1 1 105 . 100 LEU N N 117.8 0.1 1 106 . 101 ALA N N 123.8 0.1 1 107 . 102 ASP N N 115.0 0.1 1 108 . 103 VAL N N 114.9 0.1 1 109 . 104 LEU N N 120.8 0.1 1 110 . 105 GLN N N 124.7 0.1 1 111 . 105 GLN NE2 N 113.4 0.1 1 stop_ save_