data_4944 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H, 13C and 15N backbone resonance assignment of the arsenate reductase from Staphylococcus aureus in its reduced state. ; _BMRB_accession_number 4944 _BMRB_flat_file_name bmr4944.str _Entry_type original _Submission_date 2001-01-22 _Accession_date 2001-01-22 _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 Jacobs Doris M. . 2 Messens Joris . . 3 Wechselberger Rainer . . 4 Brosens Elke . . 5 Wyns Lode . . 6 Willem Rudolph . . 7 Martins Jose C. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 244 "13C chemical shifts" 358 "15N chemical shifts" 122 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2001-05-17 original author . stop_ _Original_release_date 2001-05-17 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Letter to the Editor: 1H, 13C and 15N backbone resonance assignment of the arsenate reductase from Staphylococcus aureus in its reduced state ; _Citation_status published _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 Jacobs Doris M. . 2 Messens Joris . . 3 Wechselberger Rainer W. . 4 Brosens Elke . . 5 Willem Rudolph . . 6 Wyns Lode . . 7 Martins Jose C. . stop_ _Journal_abbreviation 'J. Biomol. NMR' _Journal_volume 20 _Journal_issue 1 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 95 _Page_last 96 _Year 2001 _Details ; See references in saveframes ref_1, ref_2, ref-3, and ref_4. ; loop_ _Keyword arsenate 'arsenate reductase' metal-resistance ArsC thioredoxin stop_ save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Ji G, Silver S. Reduction of arsenate to arsenite by the ArsC protein of the arsenic resistance operon of Staphylococcus aureus plasmid pI258. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9474-8. ; _Citation_title 'Reduction of arsenate to arsenite by the ArsC protein of the arsenic resistance operon of Staphylococcus aureus plasmid pI258.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 1409657 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Ji G . . 2 Silver S . . stop_ _Journal_abbreviation 'Proc. Natl. Acad. Sci. U.S.A.' _Journal_name_full 'Proceedings of the National Academy of Sciences of the United States of America' _Journal_volume 89 _Journal_issue 20 _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 9474 _Page_last 9478 _Year 1992 _Details ; The arsenic resistance operon of Staphylococcus aureus plasmid pI258 consists of three genes, arsR (encoding the repressor regulatory protein), arsB (the determinant of the membrane efflux protein that confers resistance by pumping arsenic from the cells), and arsC (the small gene whose protein product is required for arsenate resistance only, not for arsenite resistance). ArsC has now been shown to be an arsenate reductase, converting intracellular arsenate [As(V)] to arsenite [As(III)], which is then exported from the cells by an energy-dependent efflux process. The arsenate reductase activity was found in the soluble cytoplasmic fraction in Escherichia coli (and not associated with the periplasmic fraction or the sedimentable cell envelope). Purified ArsC protein coupled in vitro with thioredoxin plus dithiothreitol (but not 2-mercaptoethanol or reduced glutathione) to reduce arsenate to arsenite. ; save_ save_ref_2 _Saveframe_category citation _Citation_full ; Ji G, Garber EA, Armes LG, Chen CM, Fuchs JA, Silver S. Arsenate reductase of Staphylococcus aureus plasmid pI258. Biochemistry. 1994 Jun 14;33(23):7294-9. ; _Citation_title 'Arsenate reductase of Staphylococcus aureus plasmid pI258.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 8003493 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Ji G . . 2 Garber 'E A' A. . 3 Armes 'L G' G. . 4 Chen 'C M' M. . 5 Fuchs 'J A' A. . 6 Silver S . . stop_ _Journal_abbreviation Biochemistry _Journal_name_full Biochemistry _Journal_volume 33 _Journal_issue 23 _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 7294 _Page_last 7299 _Year 1994 _Details ; Arsenate reductase encoded by Staphylococcus aureus arsenic-resistance plasmid pI258 was overproduced in Escherichia coli and purified. The purified enzyme reduced radioactive arsenate to arsenite when coupled to thioredoxin, thioredoxin reductase, and NADPH. NADPH oxidation coupled to arsenate reduction also required thioredoxin and thioredoxin reductase. Glutaredoxin and reduced glutathione did not stimulate arsenate reduction. NADPH oxidation showed Michaelis-Menten kinetics with a Km of 1 microM AsO4(3-) and an apparent Vmax of 200 nmol/min per mg of protein. At high substrate concentration (above 1 mM AsO4(3-), a secondary rise in the reaction rate was observed, with a Km of 2 mM and an apparent Vmax of 450 nmol/min per mg of protein. This secondary rise also occurred upon addition of phosphate or nitrate (which were not substrates for the enzyme). Arsenite (the product of the enzyme), tellurite, and antimonite [Sb(III)] were inhibitors. Selenate (but not selenite or sulfate) was a substrate for reductase-dependent NADPH oxidation, with an apparent Km of 13 mM SeO4(2-). Arsenate reductase was purified as a monomer of 14.5 kDa, consistent with the DNA sequence. Electrospray mass spectrometry showed two molecular masses of 14,810.5 and 14,436.0 Da, suggesting that 70% of the purified protein lacked the N-terminal three amino acids; HPLC coupled to electrospray mass spectroscopy of protease digest products confirmed this conclusion and verified the entire amino acid sequence. ; save_ save_ref_3 _Saveframe_category citation _Citation_full ; Messens J, Hayburn G, Brosens E, Laus G, Wyns L. Development of a downstream process for the isolation of Staphylococcus aureus arsenate reductase overproduced in Escherichia coli. J Chromatogr B Biomed Sci Appl. 2000 Jan 14;737(1-2):167-78. ; _Citation_title 'Development of a downstream process for the isolation of Staphylococcus aureus arsenate reductase overproduced in Escherichia coli.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 10681053 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Messens J . . 2 Hayburn G . . 3 Brosens E . . 4 Laus G . . 5 Wyns L . . stop_ _Journal_abbreviation 'J. Chromatogr. B Biomed. Sci. Appl.' _Journal_name_full 'Journal of chromatography. B, Biomedical sciences and applications' _Journal_volume 737 _Journal_issue 1-2 _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 167 _Page_last 178 _Year 2000 _Details ; Arsenate reductase (ArsC) encoded by Staphylococcus aureus arsenic-resistance plasmid pI258 reduces intracellular As(V) (arsenate) to the more toxic As(III) (arsenite). In order to study the structure of ArsC and to unravel biochemical and physical properties of this redox enzyme, wild type enzyme and a number of cysteine mutants were overproduced soluble in Escherichia coli. In this paper we describe a novel purification method to obtain high production levels of highly pure enzyme. A reversed-phase method was developed to separate and analyze the many different forms of ArsC. The oxidation state and the methionine oxidized forms were determined by mass spectroscopy. ; save_ save_ref_4 _Saveframe_category citation _Citation_full ; Messens J, Hayburn G, Desmyter A, Laus G, Wyns L. The essential catalytic redox couple in arsenate reductase from Staphylococcus aureus. Biochemistry. 1999 Dec 21;38(51):16857-65. ; _Citation_title 'The essential catalytic redox couple in arsenate reductase from Staphylococcus aureus.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 10606519 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Messens J . . 2 Hayburn G . . 3 Desmyter A . . 4 Laus G . . 5 Wyns L . . stop_ _Journal_abbreviation Biochemistry _Journal_name_full Biochemistry _Journal_volume 38 _Journal_issue 51 _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 16857 _Page_last 16865 _Year 1999 _Details ; Arsenate reductase (ArsC) encoded by Staphylococcus aureus arsenic-resistance plasmid pI258 reduces intracellular As(V) (arsenate) to the more toxic As(III) (arsenite), which is subsequently extruded from the cell. ArsC couples to thioredoxin, thioredoxin reductase, and NADPH to be enzymatically active. A novel purification method leads to high production levels of highly pure enzyme. A reverse phase method was introduced to systematically analyze and control the oxidation status of the enzyme. The essential cysteinyl residues and redox couple in arsenate reductase were identified by a combination of site-specific mutagenesis and endoprotease-digest mass spectroscopy analysis. The secondary structures, as determined with CD, of wild-type ArsC and its Cys mutants showed a relatively high helical content, independent of the redox status. Mutation of Cys 10, 82, and 89 led to redox-inactive enzymes. ArsC was oxidized in a single catalytic cycle and subsequently digested with endoproteinases ArgC, AspN, and GluC. From the peptide-mass profiles, cysteines 82 and 89 were identified as the redox couple of ArsC necessary to reduce arsenate to arsenite. ; save_ ################################## # Molecular system description # ################################## save_system_ArsC _Saveframe_category molecular_system _Mol_system_name 'Arsenate reductase' _Abbreviation_common ArsC _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'arsenate reductase' $ArsC stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all free' loop_ _Biological_function 'arsenate reductase' stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_ArsC _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'Arsenate reductase' _Abbreviation_common ArsC _Molecular_mass 14812 _Mol_thiol_state 'all free' _Details ; The N-terminal segment contains a sequence identical to the conserved consensus sequence in low molecular weight tyrosine phosphatase. C82 and C89 have been identified as the redox pair involved in arsenate reduction. ; ############################## # Polymer residue sequence # ############################## _Residue_count 131 _Mol_residue_sequence ; MDKKTIYFICTGNSCRSQMA EGWGKEILGEGWNVYSAGIE THGVNPKAIEAMKEVDIDIS NHTSDLIDNDILKQSDLVVT LCSDADNNCPILPPNVKKEH WGFDDPAGKEWSEFQRVRDE IKLAIEKFKLR ; loop_ _Residue_seq_code _Residue_label 1 MET 2 ASP 3 LYS 4 LYS 5 THR 6 ILE 7 TYR 8 PHE 9 ILE 10 CYS 11 THR 12 GLY 13 ASN 14 SER 15 CYS 16 ARG 17 SER 18 GLN 19 MET 20 ALA 21 GLU 22 GLY 23 TRP 24 GLY 25 LYS 26 GLU 27 ILE 28 LEU 29 GLY 30 GLU 31 GLY 32 TRP 33 ASN 34 VAL 35 TYR 36 SER 37 ALA 38 GLY 39 ILE 40 GLU 41 THR 42 HIS 43 GLY 44 VAL 45 ASN 46 PRO 47 LYS 48 ALA 49 ILE 50 GLU 51 ALA 52 MET 53 LYS 54 GLU 55 VAL 56 ASP 57 ILE 58 ASP 59 ILE 60 SER 61 ASN 62 HIS 63 THR 64 SER 65 ASP 66 LEU 67 ILE 68 ASP 69 ASN 70 ASP 71 ILE 72 LEU 73 LYS 74 GLN 75 SER 76 ASP 77 LEU 78 VAL 79 VAL 80 THR 81 LEU 82 CYS 83 SER 84 ASP 85 ALA 86 ASP 87 ASN 88 ASN 89 CYS 90 PRO 91 ILE 92 LEU 93 PRO 94 PRO 95 ASN 96 VAL 97 LYS 98 LYS 99 GLU 100 HIS 101 TRP 102 GLY 103 PHE 104 ASP 105 ASP 106 PRO 107 ALA 108 GLY 109 LYS 110 GLU 111 TRP 112 SER 113 GLU 114 PHE 115 GLN 116 ARG 117 VAL 118 ARG 119 ASP 120 GLU 121 ILE 122 LYS 123 LEU 124 ALA 125 ILE 126 GLU 127 LYS 128 PHE 129 LYS 130 LEU 131 ARG stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2014-11-16 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value PDB 1JF8 "X-ray Structure Of Reduced C10s, C15a Arsenate Reductase From Pi258" 100.00 131 98.47 98.47 2.09e-89 PDB 1JFV "X-Ray Structure Of Oxidised C10s, C15a Arsenate Reductase From Pi258" 100.00 131 96.95 96.95 2.94e-87 PDB 1LJL "Wild Type Pi258 S. Aureus Arsenate Reductase" 100.00 131 100.00 100.00 2.56e-91 PDB 1LJU "X-Ray Structure Of C15a Arsenate Reductase From Pi258 Complexed With Arsenite" 100.00 131 98.47 98.47 4.44e-89 PDB 1LK0 "Disulfide Intermediate Of C89l Arsenate Reductase From Pi258" 100.00 131 99.24 99.24 5.51e-90 PDB 1RXE "Arsc Complexed With Mnb" 100.00 131 97.71 97.71 1.43e-88 PDB 1RXI "Pi258 Arsenate Reductase (Arsc) Triple Mutant C10sC15AC82S" 100.00 131 97.71 97.71 1.43e-88 PDB 2CD7 "Staphylococcus Aureus Pi258 Arsenate Reductase (Arsc) H62q Mutant" 100.00 131 99.24 99.24 2.33e-90 PDB 2FXI "Arsenate Reductase (Arsc From Pi258) C10sC15A DOUBLE Mutant With Sulfate In Its Active Site" 100.00 131 98.47 98.47 2.09e-89 DBJ BAB43887 "arsenate reductase [Staphylococcus aureus subsp. aureus N315]" 100.00 131 100.00 100.00 2.56e-91 DBJ BAC54538 "unnamed protein product [Staphylococcus aureus]" 100.00 131 100.00 100.00 2.56e-91 DBJ BAE18778 "arsenate reductase [Staphylococcus saprophyticus subsp. saprophyticus ATCC 15305]" 100.00 131 96.95 99.24 1.53e-88 DBJ BAE92851 "arsenic reductase [Staphylococcus aureus]" 100.00 131 100.00 100.00 2.56e-91 DBJ BAG12261 "arsenate reductase [Staphylococcus aureus]" 100.00 131 100.00 100.00 2.56e-91 EMBL CAG39708 "arsenate reductase [Staphylococcus aureus subsp. aureus MRSA252]" 100.00 131 100.00 100.00 2.56e-91 GB AAA25638 "arsenate reductase [Plasmid pI258]" 100.00 131 100.00 100.00 2.56e-91 GB ACZ58845 "Arsenate reductase [Staphylococcus aureus]" 100.00 131 100.00 100.00 2.56e-91 GB ACZ68457 "Arsenate reductase [Staphylococcus aureus]" 100.00 131 100.00 100.00 2.56e-91 GB ACZ68855 "Arsenate reductase [Staphylococcus aureus]" 100.00 131 100.00 100.00 2.56e-91 GB ACZ68904 "Arsenate reductase [Staphylococcus aureus]" 100.00 131 100.00 100.00 2.56e-91 REF NP_395554 "arsenate reductase [Staphylococcus aureus subsp. aureus N315]" 100.00 131 100.00 100.00 2.56e-91 REF WP_000358995 "arsenate reductase [Staphylococcus aureus]" 100.00 131 100.00 100.00 2.56e-91 REF WP_002452591 "arsenate reductase [Staphylococcus capitis]" 61.07 80 97.50 100.00 2.54e-48 REF WP_002508683 "arsenate reductase [Staphylococcus sp. OJ82]" 100.00 131 96.95 98.47 2.26e-88 REF WP_003756625 "MULTISPECIES: arsenate reductase [Bacillales]" 100.00 131 99.24 100.00 1.57e-90 SP P0A005 "RecName: Full=Protein ArsC; AltName: Full=Arsenate reductase; AltName: Full=Arsenical pump modifier; AltName: Full=Low molecula" 100.00 131 100.00 100.00 2.56e-91 SP P0A006 "RecName: Full=Protein ArsC; AltName: Full=Arsenate reductase; AltName: Full=Arsenical pump modifier; AltName: Full=Low molecula" 100.00 131 100.00 100.00 2.56e-91 SP Q49WS7 "RecName: Full=Protein ArsC 1; AltName: Full=Arsenate reductase 1; AltName: Full=Arsenical pump modifier 1; AltName: Full=Low mo" 100.00 131 96.95 99.24 1.53e-88 SP Q6GIZ3 "RecName: Full=Protein ArsC; AltName: Full=Arsenate reductase; AltName: Full=Arsenical pump modifier; AltName: Full=Low molecula" 100.00 131 100.00 100.00 2.56e-91 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Plasmid _Gene_mnemonic $ArsC 'Staphylococcus aureus' 1280 Eubacteria . Staphylococcus aureus pI258 ars 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 $ArsC 'recombinant technology' 'E. coli' Escherichia coli BL21 plasmid pET11a 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 $ArsC 1.8 mM '[U-95% 13C; U-98% 15N]' DTT 1.0 mM . EDTA 0.1 mM . K2SO4 50 mM . 'potassium phosphate buffer' 50 mM . stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $ArsC 1.8 mM '[U-98% 15N]' DTT 1.0 mM . EDTA 0.1 mM . K2SO4 50 mM . 'potassium phosphate buffer' 50 mM . stop_ save_ ############################ # Computer software used # ############################ save_FELIX _Saveframe_category software _Name FELIX _Version 97.0 loop_ _Task processing 'manual peak picking' 'manual assignment' stop_ _Details 'commercial software from MSI (San Diege, CA)' save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model AMX _Field_strength 500 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DRX _Field_strength 600 _Details . save_ save_NMR_spectrometer_3 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model Inova _Field_strength 750 _Details . save_ ############################# # NMR applied experiments # ############################# save_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC' _Sample_label . save_ save_1H-15N_HSQC-TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC-TOCSY' _Sample_label . save_ save_1H-15N_HSQC-NOESY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC-NOESY' _Sample_label . save_ save_HNCA_4 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA _Sample_label . save_ save_HNCO_5 _Saveframe_category NMR_applied_experiment _Experiment_name HNCO _Sample_label . save_ save_HN(CO)CA_6 _Saveframe_category NMR_applied_experiment _Experiment_name HN(CO)CA _Sample_label . save_ save_HN(CA)CO_7 _Saveframe_category NMR_applied_experiment _Experiment_name HN(CA)CO _Sample_label . save_ save_CBCA(CO)NH_8 _Saveframe_category NMR_applied_experiment _Experiment_name CBCA(CO)NH _Sample_label . save_ save_HNCACB_9 _Saveframe_category NMR_applied_experiment _Experiment_name HNCACB _Sample_label . save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC-TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC-NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_4 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_5 _Saveframe_category NMR_applied_experiment _Experiment_name HNCO _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_6 _Saveframe_category NMR_applied_experiment _Experiment_name HN(CO)CA _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_7 _Saveframe_category NMR_applied_experiment _Experiment_name HN(CA)CO _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_8 _Saveframe_category NMR_applied_experiment _Experiment_name CBCA(CO)NH _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_9 _Saveframe_category NMR_applied_experiment _Experiment_name HNCACB _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_Ex-cond_1 _Saveframe_category sample_conditions _Details ; ArsC is sensitive to oxidation. Therefore all sample preparation was performed under reducing conditions, i.e. using argon flushed solutions to which DTT was added, at all times ; loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 6.7 0.1 n/a temperature 298 0.5 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 H 1 'methyl protons' ppm 0.0 internal direct . . . . DSS N 15 'methyl protons' ppm 0.0 . indirect . . . 0.101329118 DSS C 13 'methyl protons' ppm 0.0 . indirect . . . 0.251449530 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_shift_set_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '1H-15N HSQC' '1H-15N HSQC-TOCSY' '1H-15N HSQC-NOESY' HNCA HNCO HN(CO)CA HN(CA)CO CBCA(CO)NH HNCACB stop_ _Sample_conditions_label $Ex-cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'arsenate reductase' _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 MET C C 174.67 0.2 1 2 . 1 MET CA C 57.36 0.2 1 3 . 1 MET CB C 40.92 0.2 1 4 . 2 ASP H H 7.93 0.2 1 5 . 2 ASP HA H 4.38 0.02 1 6 . 2 ASP C C 176.78 0.2 1 7 . 2 ASP CA C 54.11 0.2 1 8 . 2 ASP N N 116.82 0.2 1 9 . 2 ASP CB C 38.02 0.2 1 10 . 3 LYS H H 8.29 0.2 1 11 . 3 LYS HA H 4.11 0.02 1 12 . 3 LYS C C 174.2 0.2 1 13 . 3 LYS CA C 55.52 0.2 1 14 . 3 LYS N N 122.57 0.2 1 15 . 3 LYS CB C 33.76 0.2 1 16 . 4 LYS H H 7.23 0.2 1 17 . 4 LYS HA H 4.27 0.02 1 18 . 4 LYS C C 171.62 0.2 1 19 . 4 LYS CA C 54.51 0.2 1 20 . 4 LYS N N 124.3 0.2 1 21 . 4 LYS CB C 32.75 0.2 1 22 . 5 THR H H 9.89 0.2 1 23 . 5 THR HA H 5.56 0.02 1 24 . 5 THR C C 167.17 0.2 1 25 . 5 THR CA C 62.1 0.2 1 26 . 5 THR N N 119.12 0.2 1 27 . 5 THR CB C 70.2 0.2 1 28 . 6 ILE H H 9.25 0.2 1 29 . 6 ILE HA H 5.46 0.02 1 30 . 6 ILE C C 172.79 0.2 1 31 . 6 ILE CA C 57.01 0.2 1 32 . 6 ILE N N 130.95 0.2 1 33 . 6 ILE CB C 39.83 0.2 1 34 . 7 TYR H H 8.23 0.2 1 35 . 7 TYR HA H 5.31 0.02 1 36 . 7 TYR C C 173.26 0.2 1 37 . 7 TYR CA C 49.95 0.2 1 38 . 7 TYR N N 130.71 0.2 1 39 . 7 TYR CB C 39.33 0.2 1 40 . 8 PHE H H 8.98 0.2 1 41 . 8 PHE HA H 4.98 0.02 1 42 . 8 PHE CA C 56.53 0.2 1 43 . 8 PHE N N 126.02 0.2 1 44 . 8 PHE CB C 39.83 0.2 1 45 . 9 ILE H H 9.16 0.2 1 46 . 9 ILE HA H 5.56 0.02 1 47 . 9 ILE CA C 57.04 0.2 1 48 . 9 ILE N N 122.08 0.2 1 49 . 9 ILE CB C 39.83 0.2 1 50 . 10 CYS H H 8.51 0.2 1 51 . 10 CYS HA H 4.6 0.02 1 52 . 10 CYS CA C 56.03 0.2 1 53 . 10 CYS N N 127.38 0.2 1 54 . 10 CYS CB C 28.7 0.2 1 55 . 11 THR H H 8.56 0.2 1 56 . 11 THR CA C 64.63 0.2 1 57 . 11 THR N N 117.03 0.2 1 58 . 11 THR CB C 66.66 0.2 1 59 . 12 GLY H H 8.11 0.2 1 60 . 12 GLY CA C 41.86 0.2 1 61 . 12 GLY N N 110.13 0.2 1 62 . 13 ASN H H 8.06 0.2 1 63 . 13 ASN CA C 58.56 0.2 1 64 . 13 ASN N N 118.26 0.2 1 65 . 13 ASN CB C 38.82 0.2 1 66 . 14 SER H H 8.61 0.2 1 67 . 14 SER CA C 59.07 0.2 1 68 . 14 SER N N 117.15 0.2 1 69 . 14 SER CB C 64.63 0.2 1 70 . 15 CYS H H 9.23 0.2 1 71 . 15 CYS CA C 59.07 0.2 1 72 . 15 CYS N N 120.36 0.2 1 73 . 15 CYS CB C 27.18 0.2 1 74 . 16 ARG H H 10.5 0.2 1 75 . 16 ARG CA C 59.07 0.2 1 76 . 16 ARG N N 127.13 0.2 1 77 . 16 ARG CB C 30.72 0.2 1 78 . 17 SER H H 9.61 0.2 1 79 . 17 SER CA C 59.07 0.2 1 80 . 17 SER N N 113.33 0.2 1 81 . 17 SER CB C 60.08 0.2 1 82 . 18 GLN H H 6.04 0.2 1 83 . 18 GLN HA H 4.44 0.02 1 84 . 18 GLN C C 173.26 0.2 1 85 . 18 GLN CA C 56.53 0.2 1 86 . 18 GLN N N 120.85 0.2 1 87 . 18 GLN CB C 26.17 0.2 1 88 . 19 MET H H 8.27 0.2 1 89 . 19 MET HA H 3.87 0.02 1 90 . 19 MET C C 177.48 0.2 1 91 . 19 MET CA C 59.07 0.2 1 92 . 19 MET N N 119.49 0.2 1 93 . 19 MET CB C 32.24 0.2 1 94 . 20 ALA H H 8.38 0.2 1 95 . 20 ALA HA H 3.58 0.02 1 96 . 20 ALA C C 178.42 0.2 1 97 . 20 ALA CA C 54 0.2 1 98 . 20 ALA N N 121.46 0.2 1 99 . 20 ALA CB C 15.03 0.2 1 100 . 21 GLU H H 7.83 0.2 1 101 . 21 GLU HA H 3.88 0.02 1 102 . 21 GLU C C 178.42 0.2 1 103 . 21 GLU CA C 58.05 0.2 1 104 . 21 GLU N N 119.49 0.2 1 105 . 21 GLU CB C 28.19 0.2 1 106 . 22 GLY H H 8.37 0.2 1 107 . 22 GLY HA2 H 3.83 0.02 2 108 . 22 GLY HA3 H 3.27 0.02 2 109 . 22 GLY C C 174.67 0.2 1 110 . 22 GLY CA C 46.41 0.2 1 111 . 22 GLY N N 105.69 0.2 1 112 . 23 TRP H H 8.92 0.2 1 113 . 23 TRP HA H 4.8 0.02 1 114 . 23 TRP C C 180.06 0.2 1 115 . 23 TRP CA C 58.05 0.2 1 116 . 23 TRP N N 121.71 0.2 1 117 . 23 TRP CB C 31.23 0.2 1 118 . 24 GLY H H 9.42 0.2 1 119 . 24 GLY HA2 H 3.72 0.02 2 120 . 24 GLY HA3 H 2.44 0.02 2 121 . 24 GLY C C 173.5 0.2 1 122 . 24 GLY CA C 46.5 0.2 1 123 . 24 GLY N N 109.02 0.2 1 124 . 25 LYS H H 8.36 0.2 1 125 . 25 LYS HA H 4.09 0.02 1 126 . 25 LYS C C 179.12 0.2 1 127 . 25 LYS CA C 59.07 0.2 1 128 . 25 LYS N N 122.7 0.2 1 129 . 25 LYS CB C 31.23 0.2 1 130 . 26 GLU H H 7.14 0.2 1 131 . 26 GLU HA H 4.29 0.02 1 132 . 26 GLU C C 178.19 0.2 1 133 . 26 GLU CA C 57.55 0.2 1 134 . 26 GLU N N 118.88 0.2 1 135 . 26 GLU CB C 29.2 0.2 1 136 . 27 ILE H H 8.97 0.2 1 137 . 27 ILE HA H 4.03 0.02 1 138 . 27 ILE C C 178.19 0.2 1 139 . 27 ILE CA C 62.58 0.2 1 140 . 27 ILE N N 118.38 0.2 1 141 . 27 ILE CB C 38.82 0.2 1 142 . 28 LEU H H 8.7 0.2 1 143 . 28 LEU HA H 4.43 0.02 1 144 . 28 LEU C C 181.6 0.2 1 145 . 28 LEU CA C 54 0.2 1 146 . 28 LEU N N 117.89 0.2 1 147 . 28 LEU CB C 38.82 0.2 1 148 . 29 GLY H H 7.07 0.2 1 149 . 29 GLY HA2 H 4.22 0.02 1 150 . 29 GLY HA3 H 4.22 0.02 1 151 . 29 GLY C C 173.26 0.2 1 152 . 29 GLY CA C 45.46 0.2 1 153 . 29 GLY N N 105.69 0.2 1 154 . 30 GLU H H 8.49 0.2 1 155 . 30 GLU HA H 4.21 0.02 1 156 . 30 GLU C C 177.48 0.2 1 157 . 30 GLU CA C 57.55 0.2 1 158 . 30 GLU N N 119.12 0.2 1 159 . 30 GLU CB C 28.19 0.2 1 160 . 31 GLY H H 8.42 0.2 1 161 . 31 GLY HA2 H 4.32 0.02 2 162 . 31 GLY HA3 H 3.74 0.02 2 163 . 31 GLY C C 171.39 0.2 1 164 . 31 GLY CA C 43.88 0.2 1 165 . 31 GLY N N 106.06 0.2 1 166 . 32 TRP H H 8.04 0.2 1 167 . 32 TRP HA H 5.33 0.02 1 168 . 32 TRP C C 175.14 0.2 1 169 . 32 TRP CA C 55.02 0.2 1 170 . 32 TRP N N 120.97 0.2 1 171 . 32 TRP CB C 30.22 0.2 1 172 . 33 ASN H H 9.9 0.2 1 173 . 33 ASN HA H 4.96 0.02 1 174 . 33 ASN C C 172.09 0.2 1 175 . 33 ASN CA C 51.47 0.2 1 176 . 33 ASN N N 122.2 0.2 1 177 . 33 ASN CB C 38.82 0.2 1 178 . 34 VAL H H 8.27 0.2 1 179 . 34 VAL HA H 4.68 0.02 1 180 . 34 VAL C C 173.5 0.2 1 181 . 34 VAL CA C 59.57 0.2 1 182 . 34 VAL N N 124.67 0.2 1 183 . 34 VAL CB C 32.24 0.2 1 184 . 35 TYR H H 8.38 0.2 1 185 . 35 TYR HA H 5.12 0.02 1 186 . 35 TYR C C 173.73 0.2 1 187 . 35 TYR CA C 54 0.2 1 188 . 35 TYR N N 125.41 0.2 1 189 . 35 TYR CB C 42.87 0.2 1 190 . 36 SER H H 9.4 0.2 1 191 . 36 SER HA H 6.15 0.02 1 192 . 36 SER C C 171.39 0.2 1 193 . 36 SER CA C 56.53 0.2 1 194 . 36 SER N N 116.53 0.2 1 195 . 36 SER CB C 67.67 0.2 1 196 . 37 ALA H H 8.87 0.2 1 197 . 37 ALA HA H 3.83 0.02 1 198 . 37 ALA C C 180.1 0.2 1 199 . 37 ALA CA C 50.46 0.2 1 200 . 37 ALA N N 123.07 0.2 1 201 . 37 ALA CB C 23.13 0.2 1 202 . 38 GLY H H 8.62 0.2 1 203 . 38 GLY HA2 H 4.94 0.02 2 204 . 38 GLY HA3 H 3.31 0.02 2 205 . 38 GLY CA C 41.35 0.2 1 206 . 38 GLY N N 104.83 0.2 1 207 . 39 ILE CA C 62.61 0.2 1 208 . 39 ILE CB C 37.3 0.2 1 209 . 40 GLU H H 8.13 0.2 1 210 . 40 GLU CA C 52.99 0.2 1 211 . 40 GLU N N 117.4 0.2 1 212 . 40 GLU CB C 32.24 0.2 1 213 . 41 THR H H 8 0.2 1 214 . 41 THR CA C 57.55 0.2 1 215 . 41 THR N N 109.63 0.2 1 216 . 41 THR CB C 69.19 0.2 1 217 . 43 GLY H H 7.54 0.2 1 218 . 43 GLY HA2 H 4.47 0.02 2 219 . 43 GLY HA3 H 3.85 0.02 2 220 . 43 GLY C C 170.92 0.2 1 221 . 43 GLY CA C 43.38 0.2 1 222 . 43 GLY N N 109.26 0.2 1 223 . 44 VAL H H 8.84 0.2 1 224 . 44 VAL HA H 3.82 0.02 1 225 . 44 VAL C C 175.37 0.2 1 226 . 44 VAL CA C 63.12 0.2 1 227 . 44 VAL N N 122.7 0.2 1 228 . 44 VAL CB C 31.23 0.2 1 229 . 45 ASN H H 8.62 0.2 1 230 . 45 ASN HA H 4.51 0.02 1 231 . 45 ASN C C 174.67 0.2 1 232 . 45 ASN CA C 50.46 0.2 1 233 . 45 ASN N N 131.08 0.2 1 234 . 45 ASN CB C 38.82 0.2 1 235 . 46 PRO C C 180.29 0.2 1 236 . 46 PRO CA C 64.13 0.2 1 237 . 46 PRO CB C 32.24 0.2 1 238 . 47 LYS H H 8.55 0.2 1 239 . 47 LYS HA H 3.97 0.02 1 240 . 47 LYS C C 180.06 0.2 1 241 . 47 LYS CA C 57.55 0.2 1 242 . 47 LYS N N 118.14 0.2 1 243 . 47 LYS CB C 30.72 0.2 1 244 . 48 ALA H H 7.84 0.2 1 245 . 48 ALA HA H 4.22 0.02 1 246 . 48 ALA C C 181.48 0.2 1 247 . 48 ALA CA C 53.60 0.2 1 248 . 48 ALA N N 122.57 0.2 1 249 . 48 ALA CB C 17.56 0.2 1 250 . 49 ILE H H 7.25 0.2 1 251 . 49 ILE HA H 3.29 0.02 1 252 . 49 ILE C C 175.84 0.2 1 253 . 49 ILE CA C 64.13 0.2 1 254 . 49 ILE N N 118.51 0.2 1 255 . 49 ILE CB C 36.29 0.2 1 256 . 50 GLU H H 8.09 0.2 1 257 . 50 GLU HA H 3.97 0.02 1 258 . 50 GLU C C 179.82 0.2 1 259 . 50 GLU CA C 58.05 0.2 1 260 . 50 GLU N N 122.57 0.2 1 261 . 50 GLU CB C 28.7 0.2 1 262 . 51 ALA H H 8.71 0.2 1 263 . 51 ALA HA H 4.2 0.02 1 264 . 51 ALA C C 181.7 0.2 1 265 . 51 ALA CA C 54.51 0.2 1 266 . 51 ALA N N 123.93 0.2 1 267 . 51 ALA CB C 16.04 0.2 1 268 . 52 MET H H 7.63 0.2 1 269 . 52 MET HA H 4.5 0.02 1 270 . 52 MET C C 178.89 0.2 1 271 . 52 MET CA C 53.5 0.2 1 272 . 52 MET N N 116.17 0.2 1 273 . 52 MET CB C 27.68 0.2 1 274 . 53 LYS H H 8.13 0.2 1 275 . 53 LYS HA H 3.88 0.02 1 276 . 53 LYS C C 181.47 0.2 1 277 . 53 LYS CA C 58.05 0.2 1 278 . 53 LYS N N 124.91 0.2 1 279 . 53 LYS CB C 30.68 0.2 1 280 . 54 GLU H H 7.59 0.2 1 281 . 54 GLU HA H 4.15 0.02 1 282 . 54 GLU C C 177.72 0.2 1 283 . 54 GLU CA C 58.05 0.2 1 284 . 54 GLU N N 119.25 0.2 1 285 . 54 GLU CB C 29.2 0.2 1 286 . 55 VAL H H 7.17 0.2 1 287 . 55 VAL HA H 4.61 0.02 1 288 . 55 VAL C C 171.62 0.2 1 289 . 55 VAL CA C 58.56 0.2 1 290 . 55 VAL N N 110.13 0.2 1 291 . 55 VAL CB C 28.7 0.2 1 292 . 56 ASP H H 7.78 0.2 1 293 . 56 ASP HA H 4.22 0.02 1 294 . 56 ASP C C 173.27 0.2 1 295 . 56 ASP CA C 54.51 0.2 1 296 . 56 ASP N N 115.67 0.2 1 297 . 56 ASP CB C 37.81 0.2 1 298 . 57 ILE H H 7.76 0.2 1 299 . 57 ILE HA H 4.06 0.02 1 300 . 57 ILE C C 172.56 0.2 1 301 . 57 ILE CA C 58.56 0.2 1 302 . 57 ILE N N 120.11 0.2 1 303 . 57 ILE CB C 37.81 0.2 1 304 . 58 ASP H H 8.82 0.2 1 305 . 58 ASP HA H 4.5 0.02 1 306 . 58 ASP C C 176.78 0.2 1 307 . 58 ASP CA C 51.47 0.2 1 308 . 58 ASP N N 128.49 0.2 1 309 . 58 ASP CB C 39.83 0.2 1 310 . 59 ILE H H 8.32 0.2 1 311 . 59 ILE HA H 4.16 0.02 1 312 . 59 ILE C C 175.84 0.2 1 313 . 59 ILE CA C 59.07 0.2 1 314 . 59 ILE N N 120.97 0.2 1 315 . 59 ILE CB C 35.28 0.2 1 316 . 60 SER H H 8.71 0.2 1 317 . 60 SER HA H 4.77 0.02 1 318 . 60 SER C C 173.5 0.2 1 319 . 60 SER CA C 60.58 0.2 1 320 . 60 SER N N 120.23 0.2 1 321 . 60 SER CB C 61.6 0.2 1 322 . 61 ASN H H 8.3 0.2 1 323 . 61 ASN HA H 5.05 0.02 1 324 . 61 ASN C C 175.14 0.2 1 325 . 61 ASN CA C 51.47 0.2 1 326 . 61 ASN N N 118.63 0.2 1 327 . 61 ASN CB C 37.3 0.2 1 328 . 62 HIS H H 7.63 0.2 1 329 . 62 HIS HA H 4.65 0.02 1 330 . 62 HIS C C 173.96 0.2 1 331 . 62 HIS CA C 55.02 0.2 1 332 . 62 HIS N N 121.96 0.2 1 333 . 62 HIS CB C 28.7 0.2 1 334 . 63 THR H H 8.84 0.2 1 335 . 63 THR HA H 4.43 0.02 1 336 . 63 THR C C 172.33 0.2 1 337 . 63 THR CA C 58.56 0.2 1 338 . 63 THR N N 112.34 0.2 1 339 . 63 THR CB C 68.68 0.2 1 340 . 64 SER H H 8.62 0.2 1 341 . 64 SER HA H 4.8 0.02 1 342 . 64 SER C C 171.39 0.2 1 343 . 64 SER CA C 55.02 0.2 1 344 . 64 SER N N 119.49 0.2 1 345 . 64 SER CB C 62.61 0.2 1 346 . 65 ASP H H 10.28 0.2 1 347 . 65 ASP HA H 5.09 0.02 1 348 . 65 ASP CA C 52.99 0.2 1 349 . 65 ASP N N 132.07 0.2 1 350 . 65 ASP CB C 45.4 0.2 1 351 . 66 LEU H H 8.07 0.2 1 352 . 66 LEU HA H 4.91 0.02 1 353 . 66 LEU C C 179.59 0.2 1 354 . 66 LEU CA C 52.49 0.2 1 355 . 66 LEU N N 121.96 0.2 1 356 . 66 LEU CB C 41.86 0.2 1 357 . 67 ILE H H 7.87 0.2 1 358 . 67 ILE HA H 3.34 0.02 1 359 . 67 ILE C C 173.03 0.2 1 360 . 67 ILE CA C 62.61 0.2 1 361 . 67 ILE N N 122.08 0.2 1 362 . 67 ILE CB C 36.8 0.2 1 363 . 68 ASP H H 8.32 0.2 1 364 . 68 ASP HA H 4.8 0.02 1 365 . 68 ASP C C 176.31 0.2 1 366 . 68 ASP CA C 50.97 0.2 1 367 . 68 ASP N N 130.21 0.2 1 368 . 68 ASP CB C 41.86 0.2 1 369 . 69 ASN H H 9.12 0.2 1 370 . 69 ASN HA H 4.3 0.02 1 371 . 69 ASN C C 177.25 0.2 1 372 . 69 ASN CA C 55.02 0.2 1 373 . 69 ASN N N 127.25 0.2 1 374 . 69 ASN CB C 37.81 0.2 1 375 . 70 ASP H H 8.51 0.2 1 376 . 70 ASP HA H 4.41 0.02 1 377 . 70 ASP C C 179.36 0.2 1 378 . 70 ASP CA C 56.03 0.2 1 379 . 70 ASP N N 120.35 0.2 1 380 . 70 ASP CB C 38.82 0.2 1 381 . 71 ILE H H 7.37 0.2 1 382 . 71 ILE HA H 3.35 0.02 1 383 . 71 ILE C C 181 0.2 1 384 . 71 ILE CA C 62.61 0.2 1 385 . 71 ILE N N 122.08 0.2 1 386 . 71 ILE CB C 35.28 0.2 1 387 . 72 LEU H H 8.14 0.2 1 388 . 72 LEU HA H 3.88 0.02 1 389 . 72 LEU C C 179.83 0.2 1 390 . 72 LEU CA C 57.55 0.2 1 391 . 72 LEU N N 122.94 0.2 1 392 . 72 LEU CB C 41.35 0.2 1 393 . 73 LYS H H 7.42 0.2 1 394 . 73 LYS HA H 3.98 0.02 1 395 . 73 LYS C C 177.01 0.2 1 396 . 73 LYS CA C 58.05 0.2 1 397 . 73 LYS N N 113.33 0.2 1 398 . 73 LYS CB C 31.23 0.2 1 399 . 74 GLN H H 7.18 0.2 1 400 . 74 GLN HA H 4.33 0.02 1 401 . 74 GLN C C 175.61 0.2 1 402 . 74 GLN CA C 53.5 0.2 1 403 . 74 GLN N N 117.03 0.2 1 404 . 74 GLN CB C 29.2 0.2 1 405 . 75 SER H H 7.32 0.2 1 406 . 75 SER HA H 3.94 0.02 1 407 . 75 SER C C 171.62 0.2 1 408 . 75 SER CA C 60.58 0.2 1 409 . 75 SER N N 118.38 0.2 1 410 . 75 SER CB C 62.6 0.2 1 411 . 76 ASP H H 9.12 0.2 1 412 . 76 ASP HA H 4.66 0.02 1 413 . 76 ASP C C 175.84 0.2 1 414 . 76 ASP CA C 55.52 0.2 1 415 . 76 ASP N N 123.81 0.2 1 416 . 76 ASP CB C 41.35 0.2 1 417 . 77 LEU H H 7.8 0.2 1 418 . 77 LEU HA H 4.41 0.02 1 419 . 77 LEU C C 171.16 0.2 1 420 . 77 LEU CA C 53.5 0.2 1 421 . 77 LEU N N 121.59 0.2 1 422 . 77 LEU CB C 44.39 0.2 1 423 . 78 VAL H H 8.82 0.2 1 424 . 78 VAL HA H 4.71 0.02 1 425 . 78 VAL C C 172.09 0.2 1 426 . 78 VAL CA C 59.07 0.2 1 427 . 78 VAL N N 128.49 0.2 1 428 . 78 VAL CB C 32.24 0.2 1 429 . 79 VAL H H 8.42 0.2 1 430 . 79 VAL HA H 4.7 0.02 1 431 . 79 VAL C C 176.08 0.2 1 432 . 79 VAL CA C 58.05 0.2 1 433 . 79 VAL N N 127.74 0.2 1 434 . 79 VAL CB C 32.75 0.2 1 435 . 80 THR H H 8.9 0.2 1 436 . 80 THR HA H 5.31 0.02 1 437 . 80 THR C C 175.84 0.2 1 438 . 80 THR CA C 57.55 0.2 1 439 . 80 THR N N 117.89 0.2 1 440 . 80 THR CB C 68.18 0.2 1 441 . 81 LEU H H 7.95 0.2 1 442 . 81 LEU HA H 5.8 0.02 1 443 . 81 LEU CA C 55.02 0.2 1 444 . 81 LEU N N 121.59 0.2 1 445 . 81 LEU CB C 43.38 0.2 1 446 . 82 CYS H H 7.63 0.2 1 447 . 82 CYS HA H 4.46 0.02 1 448 . 82 CYS C C 171.86 0.2 1 449 . 82 CYS CA C 56.03 0.2 1 450 . 82 CYS N N 107.54 0.2 1 451 . 82 CYS CB C 29.71 0.2 1 452 . 83 SER HA H 4.54 0.02 1 453 . 83 SER C C 175.61 0.2 1 454 . 83 SER CA C 61.6 0.2 1 455 . 83 SER CB C 61.6 0.2 1 456 . 84 ASP H H 8.74 0.2 1 457 . 84 ASP HA H 4.4 0.02 1 458 . 84 ASP C C 176.08 0.2 1 459 . 84 ASP CA C 56.53 0.2 1 460 . 84 ASP N N 122.7 0.2 1 461 . 84 ASP CB C 39.83 0.2 1 462 . 85 ALA H H 7.58 0.2 1 463 . 85 ALA HA H 4.22 0.02 1 464 . 85 ALA C C 177.95 0.2 1 465 . 85 ALA CA C 53.5 0.2 1 466 . 85 ALA N N 122.08 0.2 1 467 . 85 ALA CB C 17.56 0.2 1 468 . 86 ASP H H 7.91 0.2 1 469 . 86 ASP HA H 4.38 0.02 1 470 . 86 ASP C C 175.84 0.2 1 471 . 86 ASP CA C 57.04 0.2 1 472 . 86 ASP N N 118.38 0.2 1 473 . 86 ASP CB C 40.84 0.2 1 474 . 87 ASN H H 8.29 0.2 1 475 . 87 ASN HA H 4.44 0.02 1 476 . 87 ASN C C 173.97 0.2 1 477 . 87 ASN CA C 54 0.2 1 478 . 87 ASN N N 116.29 0.2 1 479 . 87 ASN CB C 37.3 0.2 1 480 . 88 ASN H H 7.51 0.2 1 481 . 88 ASN HA H 4.61 0.02 1 482 . 88 ASN C C 175.37 0.2 1 483 . 88 ASN CA C 52.07 0.2 1 484 . 88 ASN N N 116.78 0.2 1 485 . 88 ASN CB C 38.31 0.2 1 486 . 89 CYS H H 7.16 0.2 1 487 . 89 CYS HA H 4.9 0.02 1 488 . 89 CYS CA C 56.53 0.2 1 489 . 89 CYS N N 123.93 0.2 1 490 . 89 CYS CB C 26.17 0.2 1 491 . 90 PRO C C 173.5 0.2 1 492 . 90 PRO CA C 61.6 0.2 1 493 . 90 PRO CB C 31.23 0.2 1 494 . 91 ILE H H 7.99 0.2 1 495 . 91 ILE HA H 3.99 0.02 1 496 . 91 ILE C C 175.37 0.2 1 497 . 91 ILE CA C 58.59 0.2 1 498 . 91 ILE N N 121.34 0.2 1 499 . 91 ILE CB C 34.09 0.2 1 500 . 92 LEU H H 8.28 0.2 1 501 . 92 LEU HA H 4.68 0.02 1 502 . 92 LEU C C 173.03 0.2 1 503 . 92 LEU CA C 50.97 0.2 1 504 . 92 LEU N N 129.47 0.2 1 505 . 92 LEU CB C 40.33 0.2 1 506 . 94 PRO C C 176.31 0.2 1 507 . 94 PRO CA C 63.12 0.2 1 508 . 94 PRO CB C 31.23 0.2 1 509 . 95 ASN H H 8.36 0.2 1 510 . 95 ASN HA H 4.55 0.02 1 511 . 95 ASN C C 173.5 0.2 1 512 . 95 ASN CA C 52.99 0.2 1 513 . 95 ASN N N 113.58 0.2 1 514 . 95 ASN CB C 36.29 0.2 1 515 . 96 VAL H H 7.42 0.2 1 516 . 96 VAL HA H 4.02 0.02 1 517 . 96 VAL C C 173.97 0.2 1 518 . 96 VAL CA C 62.1 0.2 1 519 . 96 VAL N N 123.44 0.2 1 520 . 96 VAL CB C 31.73 0.2 1 521 . 97 LYS H H 7.56 0.2 1 522 . 97 LYS HA H 4.27 0.02 1 523 . 97 LYS C C 173.5 0.2 1 524 . 97 LYS CA C 54.51 0.2 1 525 . 97 LYS N N 128.86 0.2 1 526 . 97 LYS CB C 32.24 0.2 1 527 . 98 LYS H H 8.29 0.2 1 528 . 98 LYS HA H 6.03 0.02 1 529 . 98 LYS C C 175.37 0.2 1 530 . 98 LYS CA C 52.99 0.2 1 531 . 98 LYS N N 123.56 0.2 1 532 . 98 LYS CB C 33.76 0.2 1 533 . 99 GLU H H 8.96 0.2 1 534 . 99 GLU HA H 4.5 0.02 1 535 . 99 GLU C C 171.62 0.2 1 536 . 99 GLU CA C 54 0.2 1 537 . 99 GLU N N 126.02 0.2 1 538 . 99 GLU CB C 34.27 0.2 1 539 . 100 HIS H H 8.6 0.2 1 540 . 100 HIS HA H 5.38 0.02 1 541 . 100 HIS C C 172.8 0.2 1 542 . 100 HIS CA C 52.49 0.2 1 543 . 100 HIS N N 121.09 0.2 1 544 . 100 HIS CB C 29.2 0.2 1 545 . 101 TRP H H 8.41 0.2 1 546 . 101 TRP HA H 4.85 0.02 1 547 . 101 TRP CA C 50.97 0.2 1 548 . 101 TRP N N 132.31 0.2 1 549 . 101 TRP CB C 29.2 0.2 1 550 . 102 GLY H H 7.35 0.2 1 551 . 102 GLY HA2 H 3.77 0.02 2 552 . 102 GLY HA3 H 3.16 0.02 2 553 . 102 GLY CA C 45.4 0.2 1 554 . 102 GLY N N 106.06 0.2 1 555 . 103 PHE H H 8.04 0.2 1 556 . 103 PHE HA H 4.7 0.02 1 557 . 103 PHE C C 173.03 0.2 1 558 . 103 PHE CA C 56.53 0.2 1 559 . 103 PHE N N 121.09 0.2 1 560 . 103 PHE CB C 42.36 0.2 1 561 . 104 ASP H H 9.4 0.2 1 562 . 104 ASP HA H 4.32 0.02 1 563 . 104 ASP C C 176.78 0.2 1 564 . 104 ASP CA C 52.99 0.2 1 565 . 104 ASP N N 127.13 0.2 1 566 . 104 ASP CB C 39.83 0.2 1 567 . 105 ASP H H 8.51 0.2 1 568 . 105 ASP HA H 4.31 0.02 1 569 . 105 ASP CA C 49.45 0.2 1 570 . 105 ASP N N 119.61 0.2 1 571 . 105 ASP CB C 40.84 0.2 1 572 . 106 PRO C C 181 0.2 1 573 . 106 PRO CA C 61.6 0.2 1 574 . 106 PRO CB C 29.2 0.2 1 575 . 107 ALA H H 7.65 0.2 1 576 . 107 ALA HA H 4.24 0.02 1 577 . 107 ALA C C 172.09 0.2 1 578 . 107 ALA CA C 53.5 0.2 1 579 . 107 ALA N N 122.2 0.2 1 580 . 107 ALA CB C 17.56 0.2 1 581 . 108 GLY H H 8.92 0.2 1 582 . 108 GLY HA2 H 4.1 0.02 2 583 . 108 GLY HA3 H 3.85 0.02 2 584 . 108 GLY C C 171.39 0.2 1 585 . 108 GLY CA C 44.39 0.2 1 586 . 108 GLY N N 111.36 0.2 1 587 . 109 LYS H H 8.16 0.2 1 588 . 109 LYS HA H 4.64 0.02 1 589 . 109 LYS C C 174.91 0.2 1 590 . 109 LYS CA C 52.99 0.2 1 591 . 109 LYS N N 121.09 0.2 1 592 . 109 LYS CB C 32.24 0.2 1 593 . 110 GLU H H 8.58 0.2 1 594 . 110 GLU HA H 4.39 0.02 1 595 . 110 GLU C C 177.01 0.2 1 596 . 110 GLU CA C 55.02 0.2 1 597 . 110 GLU N N 118.38 0.2 1 598 . 110 GLU CB C 29.2 0.2 1 599 . 111 TRP H H 8.96 0.2 1 600 . 111 TRP HA H 4.8 0.02 1 601 . 111 TRP C C 177.48 0.2 1 602 . 111 TRP CA C 58.56 0.2 1 603 . 111 TRP N N 124.42 0.2 1 604 . 111 TRP CB C 29.2 0.2 1 605 . 112 SER H H 8.71 0.2 1 606 . 112 SER HA H 4.43 0.02 1 607 . 112 SER C C 177.95 0.2 1 608 . 112 SER CA C 59.57 0.2 1 609 . 112 SER N N 112.47 0.2 1 610 . 112 SER CB C 61.09 0.2 1 611 . 113 GLU H H 8.06 0.2 1 612 . 113 GLU HA H 4.5 0.02 1 613 . 113 GLU C C 179.36 0.2 1 614 . 113 GLU CA C 56.53 0.2 1 615 . 113 GLU N N 123.81 0.2 1 616 . 113 GLU CB C 28.7 0.2 1 617 . 114 PHE H H 7.41 0.2 1 618 . 114 PHE HA H 3.45 0.02 1 619 . 114 PHE C C 178.65 0.2 1 620 . 114 PHE CA C 62.1 0.2 1 621 . 114 PHE N N 120.72 0.2 1 622 . 114 PHE CB C 37.81 0.2 1 623 . 115 GLN H H 7.26 0.2 1 624 . 115 GLN HA H 3.2 0.02 1 625 . 115 GLN C C 175.84 0.2 1 626 . 115 GLN CA C 58.05 0.2 1 627 . 115 GLN N N 116.16 0.2 1 628 . 115 GLN CB C 28.19 0.2 1 629 . 116 ARG H H 8.01 0.2 1 630 . 116 ARG HA H 3.81 0.02 1 631 . 116 ARG C C 179.83 0.2 1 632 . 116 ARG CA C 58.56 0.2 1 633 . 116 ARG N N 118.88 0.2 1 634 . 116 ARG CB C 28.7 0.2 1 635 . 117 VAL H H 7.93 0.2 1 636 . 117 VAL HA H 4.19 0.02 1 637 . 117 VAL C C 177.01 0.2 1 638 . 117 VAL CA C 66.15 0.2 1 639 . 117 VAL N N 119.98 0.2 1 640 . 117 VAL CB C 30.72 0.2 1 641 . 118 ARG H H 7.85 0.2 1 642 . 118 ARG HA H 4.06 0.02 1 643 . 118 ARG C C 180.29 0.2 1 644 . 118 ARG CA C 59.57 0.2 1 645 . 118 ARG N N 120.23 0.2 1 646 . 118 ARG CB C 30.2 0.2 1 647 . 119 ASP H H 8.02 0.2 1 648 . 119 ASP HA H 4.6 0.02 1 649 . 119 ASP C C 179.83 0.2 1 650 . 119 ASP CA C 56.03 0.2 1 651 . 119 ASP N N 122.45 0.2 1 652 . 119 ASP CB C 38.31 0.2 1 653 . 120 GLU H H 8.6 0.2 1 654 . 120 GLU HA H 3.98 0.02 1 655 . 120 GLU C C 181.7 0.2 1 656 . 120 GLU CA C 58.05 0.2 1 657 . 120 GLU N N 126.02 0.2 1 658 . 120 GLU CB C 28.7 0.2 1 659 . 121 ILE H H 8.15 0.2 1 660 . 121 ILE HA H 3.3 0.02 1 661 . 121 ILE C C 176.54 0.2 1 662 . 121 ILE CA C 64.63 0.2 1 663 . 121 ILE N N 123.93 0.2 1 664 . 121 ILE CB C 35.78 0.2 1 665 . 122 LYS H H 7.2 0.2 1 666 . 122 LYS HA H 3.87 0.02 1 667 . 122 LYS C C 177.48 0.2 1 668 . 122 LYS CA C 59.07 0.2 1 669 . 122 LYS N N 121.22 0.2 1 670 . 122 LYS CB C 31.23 0.2 1 671 . 123 LEU H H 7.17 0.2 1 672 . 123 LEU HA H 3.91 0.02 1 673 . 123 LEU C C 179.36 0.2 1 674 . 123 LEU CA C 56.02 0.2 1 675 . 123 LEU N N 117.77 0.2 1 676 . 123 LEU CB C 40.34 0.2 1 677 . 124 ALA H H 7.61 0.2 1 678 . 124 ALA HA H 3.78 0.02 1 679 . 124 ALA C C 182.4 0.2 1 680 . 124 ALA CA C 53.5 0.2 1 681 . 124 ALA N N 121.34 0.2 1 682 . 124 ALA CB C 17.06 0.2 1 683 . 125 ILE H H 7.99 0.2 1 684 . 125 ILE HA H 3.76 0.02 1 685 . 125 ILE C C 177.24 0.2 1 686 . 125 ILE CA C 64.13 0.2 1 687 . 125 ILE N N 121.83 0.2 1 688 . 125 ILE CB C 35.78 0.2 1 689 . 126 GLU H H 8.66 0.2 1 690 . 126 GLU HA H 3.81 0.02 1 691 . 126 GLU C C 181 0.2 1 692 . 126 GLU CA C 59.07 0.2 1 693 . 126 GLU N N 122.57 0.2 1 694 . 126 GLU CB C 28.19 0.2 1 695 . 127 LYS H H 8.19 0.2 1 696 . 127 LYS HA H 3.86 0.02 1 697 . 127 LYS C C 181 0.2 1 698 . 127 LYS CA C 58.05 0.2 1 699 . 127 LYS N N 118.51 0.2 1 700 . 127 LYS CB C 30.72 0.2 1 701 . 128 PHE H H 7.79 0.2 1 702 . 128 PHE HA H 4.04 0.02 1 703 . 128 PHE C C 177.48 0.2 1 704 . 128 PHE CA C 60.08 0.2 1 705 . 128 PHE N N 122.94 0.2 1 706 . 128 PHE CB C 38.31 0.2 1 707 . 129 LYS H H 8.04 0.2 1 708 . 129 LYS HA H 4.42 0.02 1 709 . 129 LYS C C 176.55 0.2 1 710 . 129 LYS CA C 57.55 0.2 1 711 . 129 LYS N N 120.97 0.2 1 712 . 129 LYS CB C 31.73 0.2 1 713 . 130 LEU H H 7.04 0.2 1 714 . 130 LEU HA H 4.23 0.02 1 715 . 130 LEU C C 176.55 0.2 1 716 . 130 LEU CA C 53.5 0.2 1 717 . 130 LEU N N 117.4 0.2 1 718 . 130 LEU CB C 41.35 0.2 1 719 . 131 ARG H H 7.17 0.2 1 720 . 131 ARG HA H 3.79 0.02 1 721 . 131 ARG C C 184.51 0.2 1 722 . 131 ARG CA C 58.56 0.2 1 723 . 131 ARG N N 126.39 0.2 1 724 . 131 ARG CB C 29.71 0.2 1 stop_ save_