data_4073 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Backbone 1H, 13C and 15N Chemical Shift Assignments for Oxidized Human Ferredoxin ; _BMRB_accession_number 4073 _BMRB_flat_file_name bmr4073.str _Entry_type original _Submission_date 1997-11-20 _Accession_date 1997-11-20 _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 Xia Bin . . 2 Volkman Brian F. . 3 Markley John L. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 185 "13C chemical shifts" 264 "15N chemical shifts" 89 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 1997-11-20 original author 'original release' 2002-04-08 update BMRB 'update of relationships' stop_ loop_ _Related_BMRB_accession_number _Relationship 4073 'Chemical Shift of Oxidized Human Ferredoxin' 5337 'Complete assignments of 1H, 13C and 15N Chemical Shifts for Oxidized Human Adrenodoxin (4-114)' stop_ _Original_release_date 2016-09-15 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full ; Xia, B., Volkman, B. F. , and Markley, J. L., "Evidence for Oxidation-State-Dependent Conformational Changes in Human Ferredoxin from Multinuclear, Multidimensional NMR Spectroscopy," Biochemistry 37, 3965-3973 (1998). ; _Citation_title ; Evidence for Oxidation-State-Dependent Conformational Changes in Human Ferredoxin from Multinuclear, Multidimensional NMR Spectroscopy ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code 98191359 _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Xia Bin . . 2 Volkman Brian F. . 3 Markley John L. . stop_ _Journal_abbreviation Biochemistry _Journal_volume 37 _Journal_issue . _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 3965 _Page_last 3973 _Year 1998 _Details . loop_ _Keyword 'F2S2 cluster' 'HuFd reduced' NMR adrenodoxin 'human ferrodoxin (HuFd)' stop_ save_ ####################################### # Cited references within the entry # ####################################### save_citation_one _Saveframe_category citation _Citation_full ; Xia, B., Cheng, H., Skjeldal, L., Coghlan, V. M., Vickery, L. E., and Markley, J. L., Biochemistry 34, 180-187, (1995). ; _Citation_title ; Multinuclear magnetic resonance and mutagenesis studies of the histidine residues of human mitochondrial ferredoxin. ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 7819194 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Xia B . . 2 Cheng H . . 3 Skjeldal L . . 4 Coghlan 'V M' M. . 5 Vickery 'L E' E. . 6 Markley 'J L' L. . stop_ _Journal_abbreviation Biochemistry _Journal_name_full Biochemistry _Journal_volume 34 _Journal_issue 1 _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 180 _Page_last 187 _Year 1995 _Details ; Human mitochondrial ferredoxin is a [2Fe-2S] protein that functions to transfer electrons from NADPH-dependent ferredoxin reductase to cytochrome P450 enzymes. Two of the three histidines of human ferredoxin are strictly conserved in the sequences of all known vertebrate ferredoxins, and one of these (His56) is adjacent to Cys55, which serves as one of the ligands to the iron-sulfur cluster. All but 16 of its residues show sequence identity with those of bovine ferredoxin. It has been proposed for bovine ferredoxin that His56 hydrogen bonds with a labile sulfur and that the reduction of the iron-sulfur center is accompanied by the uptake of a proton by this histidine [Lambeth, J. D., Seybert, D. W., Lancaster, J. R., Jr., Salerno, J. C., & Kamin, H. (1982) Mol. Cell. Biochem. 45, 13-31]. In this paper, we report procedures for labeling human ferredoxin uniformly with 15N using 15NH4Cl and selectively with 13C by the incorporation of [U-13C]histidine. Most of the imidazole 1H, 13C, and 15N resonances of the three histidines have been assigned by heteronuclear two-dimensional single- and multiple-bond correlation spectroscopy. Site-directed mutagenesis was used in assigning the NMR signals from His56. The pKa values of His10 (6.5) and His62 (5.8) in oxidized human ferredoxin were found to be similar to those reported previously for the corresponding residues of bovine ferredoxin [Greenfield, N. J., Wu, X., & Jordan, F. (1989) Biochim. Biophys. Acta 995, 246-254; Miura, S., Tamita, S., & Ichikawa, Y. (1991) J. Biol. Chem. 266, 19212-19216].(ABSTRACT TRUNCATED AT 250 WORDS) ; save_ save_citation_two _Saveframe_category citation _Citation_full ; Xia B., Cheng H., Bandarian V., Reed G. H., & Markley J. L., Biochemistry 35, 9488-9495,(1996). ; _Citation_title ; Human ferredoxin: overproduction in Escherichia coli, reconstitution in vitro, and spectroscopic studies of iron-sulfur cluster ligand cysteine-to-serine mutants. ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 8755728 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Xia B. . . 2 Cheng H. . . 3 Bandarian V. . . 4 Reed G.H. H. . 5 Markley J.L. L. . stop_ _Journal_abbreviation Biochemistry _Journal_name_full Biochemistry _Journal_volume 35 _Journal_issue 29 _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 9488 _Page_last 9495 _Year 1996 _Details ; Human ferredoxin, the human equivalent of bovine adrenodoxin, is a small iron-sulfur protein with one [2Fe-2S] cluster. It functions, as do other vertebrate ferredoxins, to transfer electrons during the processes of steroid hormone synthesis. A DNA fragment encoding the mature form of human ferredoxin was cloned into an expression vector under control of the T7 RNA polymerase/promoter system. The protein was overproduced in Escherichia coli, and the [2Fe-2S] cluster was incorporated into the protein by in vitro reconstitution. The overall yield was approximately 30 mg of purified, reconstituted ferredoxin per liter of culture. Four of the five cysteines in human ferredoxin are coordinated to the iron-sulfur cluster. First, the non-ligand cysteine (cysteine-95) was mutated to alanine, and then double mutants were created in which each of the other four cysteines (at positions 46, 52, 55, and 92) were mutated individually to serine. The wild-type ferredoxin and each of the five mutant proteins were studied by UV-visible spectroscopy and electron paramagnetic resonance spectroscopy. The EPR gav values of all five mutants were very similar to that of wild-type human ferredoxin. In the reduced state, three of the cysteine-to-serine mutants exhibited axial EPR spectra similar to that of wild-type, but one of the double mutants (C52S/C95A) exhibited a rhombic EPR spectrum. The UV-visible spectroscopic properties of the wild-type and the C95A mutant ferredoxins were identical, but those of the other cysteine-to-serine mutant proteins of human ferredoxin were quite different from those of the wild-type protein and each other. These results, along with those from cysteine-to-serine mutations in other ferredoxins, provide the basis for a more comprehensive theoretical and practical understanding of the features important to the ligation of [2Fe-2S] clusters, although they do not yet permit determination of which two cysteines ligate Fe(II) and which ligate Fe(III) in the reduced protein. ; save_ save_citation_three _Saveframe_category citation _Citation_full 'Chylla, R. A. & Markley, J. L. J. Magn. Reson. (Series B) 102, 148-154, (1993)' _Citation_title . _Citation_status . _Citation_type . _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Chylla R. . . 2 Markley J.L. L. . stop_ _Journal_abbreviation . _Journal_name_full . _Journal_volume . _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 . _Page_last . _Year . _Details . save_ ################################## # Molecular system description # ################################## save_system_HuFd-ox _Saveframe_category molecular_system _Mol_system_name 'oxidized human ferredoxin' _Abbreviation_common HuFd-ox _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label HuFd-ox $HuFd-ox 2Fe2S $FE stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic yes _System_thiol_state . loop_ _Biological_function 'cholesterol biosynthesis' 'electron transfer' stop_ _Database_query_date . _Details 'oxidized form of the protein' save_ ######################## # Monomeric polymers # ######################## save_HuFd-ox _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'human ferredoxin' _Name_variant HuFd-ox _Abbreviation_common HuFd _Molecular_mass 13800 _Mol_thiol_state . _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 124 _Mol_residue_sequence ; SSSEDKITVHFINRDGETLT TKGKVGDSLLDVVVENNLDI DGFGACEGTLACSTCHLIFE DHIYEKLDAITDEENDMLDL AYGLTDRSRLGCQICLTKSM DNMTVRVPETVADARQSIDV GKTS ; loop_ _Residue_seq_code _Residue_label 1 SER 2 SER 3 SER 4 GLU 5 ASP 6 LYS 7 ILE 8 THR 9 VAL 10 HIS 11 PHE 12 ILE 13 ASN 14 ARG 15 ASP 16 GLY 17 GLU 18 THR 19 LEU 20 THR 21 THR 22 LYS 23 GLY 24 LYS 25 VAL 26 GLY 27 ASP 28 SER 29 LEU 30 LEU 31 ASP 32 VAL 33 VAL 34 VAL 35 GLU 36 ASN 37 ASN 38 LEU 39 ASP 40 ILE 41 ASP 42 GLY 43 PHE 44 GLY 45 ALA 46 CYS 47 GLU 48 GLY 49 THR 50 LEU 51 ALA 52 CYS 53 SER 54 THR 55 CYS 56 HIS 57 LEU 58 ILE 59 PHE 60 GLU 61 ASP 62 HIS 63 ILE 64 TYR 65 GLU 66 LYS 67 LEU 68 ASP 69 ALA 70 ILE 71 THR 72 ASP 73 GLU 74 GLU 75 ASN 76 ASP 77 MET 78 LEU 79 ASP 80 LEU 81 ALA 82 TYR 83 GLY 84 LEU 85 THR 86 ASP 87 ARG 88 SER 89 ARG 90 LEU 91 GLY 92 CYS 93 GLN 94 ILE 95 CYS 96 LEU 97 THR 98 LYS 99 SER 100 MET 101 ASP 102 ASN 103 MET 104 THR 105 VAL 106 ARG 107 VAL 108 PRO 109 GLU 110 THR 111 VAL 112 ALA 113 ASP 114 ALA 115 ARG 116 GLN 117 SER 118 ILE 119 ASP 120 VAL 121 GLY 122 LYS 123 THR 124 SER stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ ############# # Ligands # ############# save_FE _Saveframe_category ligand _Mol_type non-polymer _Name_common "FE (FE (III) ION)" _BMRB_code . _PDB_code FE _Molecular_mass 55.845 _Mol_charge 3 _Mol_paramagnetic . _Mol_aromatic no _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Mon Jul 11 10:30:25 2011 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons FE FE FE . 3 . ? stop_ _Mol_thiol_state . _Sequence_homology_query_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $HuFd-ox Human 9609 . . Homo sapiens stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Variant _Vector_type _Vector_name $HuFd-ox 'recombinant technology' 'E. coli' Escherichia coli BL21 (DE3)pLysS plasmid HuFd/pET9a stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_one _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $HuFd-ox . mM 1 2 '[U-99.8% 15N]' Tris-HCl 50 mM . . . stop_ save_ save_sample_two _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $HuFd-ox . mM 1 2 '[U-99% 13C; U-99.8% 15N]' Tris-HCl 50 mM . . . stop_ save_ ############################ # Computer software used # ############################ save_software_Felix _Saveframe_category software _Name FELIX _Version '2.30 & 95.0' loop_ _Vendor _Address _Electronic_address 'Molecular Simulations, San Diego, CA' . . stop_ loop_ _Task 'used for all FT of data' stop_ _Details . save_ save_software_Peakpick _Saveframe_category software _Name Peakpick _Version 1.1 loop_ _Vendor _Address _Electronic_address 'Molecular Simulations, San Diego, CA' . . stop_ loop_ _Task 'used for peakpicking all data' stop_ _Details . _Citation_label $citation_three save_ save_software_PPFLX _Saveframe_category software _Name PPFLX _Version 1.1 loop_ _Vendor _Address _Electronic_address 'Molecular Simulations, San Diego, CA' . . stop_ loop_ _Task 'used for peakpicking all data' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_one _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DMX500 _Field_strength 500 _Details . save_ save_spectrometer_two _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DMX600 _Field_strength 600 _Details . save_ save_spectrometer_three _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DMX750 _Field_strength 750 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name 2D_1H-15N_HSQC _Sample_label . save_ save_3D_HNCO_2 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_HNCO _Sample_label . save_ save_3D_HNCA_3 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_HNCA _Sample_label . save_ save_3D_HN(CO)CA_4 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_HN(CO)CA _Sample_label . save_ save_3D_HNCACB_5 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_HNCACB _Sample_label . save_ save_3D_CBCA(CO)NH_6 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_CBCA(CO)NH _Sample_label . save_ save_3D_HCACO_7 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_HCACO _Sample_label . save_ save_3D_15N-TOCSYd_8 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_15N-TOCSYd _Sample_label . save_ save_3D_15N-NOESYd_9 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_15N-NOESYd _Sample_label . save_ save_3D_15N-TOCSYe_10 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_15N-TOCSYe _Sample_label . save_ save_3D_15N-NOESY_11 _Saveframe_category NMR_applied_experiment _Experiment_name 3D_15N-NOESY _Sample_label . save_ ####################### # Sample conditions # ####################### save_sample_conditions_HuFd-ox _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 7.4 0.1 pH temperature 295 1 K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chem_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 DSS C 13 'methyl protons' ppm 0.00 external indirect . . . DSS H 1 'methyl protons' ppm 0.00 internal direct . . . DSS N 15 'methyl protons' ppm 0.00 external indirect . . . 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_shift_assignment _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label 2D_1H-15N_HSQC stop_ loop_ _Sample_label $sample_one stop_ _Sample_conditions_label $sample_conditions_HuFd-ox _Chem_shift_reference_set_label $chem_shift_reference _Mol_system_component_name HuFd-ox _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 SER CA C 58.51 . 1 2 . 2 SER CB C 64.15 . 1 3 . 3 SER H H 8.63 . 1 4 . 3 SER HA H 4.44 . 1 5 . 3 SER C C 174.84 . 1 6 . 3 SER CA C 59.01 . 1 7 . 3 SER CB C 63.74 . 1 8 . 3 SER N N 118.62 . 1 9 . 4 GLU H H 8.44 . 1 10 . 4 GLU HA H 4.31 . 1 11 . 4 GLU C C 176.15 . 1 12 . 4 GLU CA C 56.71 . 1 13 . 4 GLU CB C 30.33 . 1 14 . 4 GLU N N 122.58 . 1 15 . 5 ASP H H 8.32 . 1 16 . 5 ASP HA H 4.61 . 1 17 . 5 ASP C C 175.68 . 1 18 . 5 ASP CA C 54.59 . 1 19 . 5 ASP CB C 41.13 . 1 20 . 5 ASP N N 121.84 . 1 21 . 6 LYS H H 8.11 . 1 22 . 6 LYS HA H 4.88 . 1 23 . 6 LYS C C 176.27 . 1 24 . 6 LYS CA C 55.49 . 1 25 . 6 LYS CB C 35.55 . 1 26 . 6 LYS N N 120.62 . 1 27 . 7 ILE H H 9.03 . 1 28 . 7 ILE HA H 4.47 . 1 29 . 7 ILE C C 175.02 . 1 30 . 7 ILE CA C 59.02 . 1 31 . 7 ILE CB C 40.92 . 1 32 . 7 ILE N N 119.32 . 1 33 . 8 THR H H 8.90 . 1 34 . 8 THR HA H 4.73 . 1 35 . 8 THR C C 174.18 . 1 36 . 8 THR CA C 63.49 . 1 37 . 8 THR CB C 69.17 . 1 38 . 8 THR N N 122.61 . 1 39 . 9 VAL H H 8.56 . 1 40 . 9 VAL HA H 4.21 . 1 41 . 9 VAL C C 173.52 . 1 42 . 9 VAL CA C 61.15 . 1 43 . 9 VAL CB C 35.72 . 1 44 . 9 VAL N N 125.50 . 1 45 . 10 HIS H H 8.65 . 1 46 . 10 HIS HA H 5.41 . 1 47 . 10 HIS C C 174.61 . 1 48 . 10 HIS CA C 54.22 . 1 49 . 10 HIS CB C 31.18 . 1 50 . 10 HIS N N 126.08 . 1 51 . 11 PHE H H 9.65 . 1 52 . 11 PHE HA H 5.18 . 1 53 . 11 PHE C C 175.63 . 1 54 . 11 PHE CA C 55.68 . 1 55 . 11 PHE CB C 41.89 . 1 56 . 11 PHE N N 123.24 . 1 57 . 12 ILE H H 9.11 . 1 58 . 12 ILE HA H 4.72 . 1 59 . 12 ILE C C 176.24 . 1 60 . 12 ILE CA C 60.47 . 1 61 . 12 ILE CB C 37.14 . 1 62 . 12 ILE N N 123.60 . 1 63 . 13 ASN H H 8.98 . 1 64 . 13 ASN HA H 4.68 . 1 65 . 13 ASN C C 177.29 . 1 66 . 13 ASN CA C 51.27 . 1 67 . 13 ASN CB C 38.93 . 1 68 . 13 ASN N N 125.92 . 1 69 . 14 ARG H H 8.67 . 1 70 . 14 ARG HA H 4.05 . 1 71 . 14 ARG C C 176.30 . 1 72 . 14 ARG CA C 59.33 . 1 73 . 14 ARG CB C 29.56 . 1 74 . 14 ARG N N 119.96 . 1 75 . 15 ASP H H 7.72 . 1 76 . 15 ASP HA H 4.61 . 1 77 . 15 ASP C C 177.02 . 1 78 . 15 ASP CA C 53.57 . 1 79 . 15 ASP CB C 40.70 . 1 80 . 15 ASP N N 116.90 . 1 81 . 16 GLY H H 8.22 . 1 82 . 16 GLY HA2 H 4.24 . 2 83 . 16 GLY HA3 H 3.52 . 2 84 . 16 GLY C C 174.39 . 1 85 . 16 GLY CA C 45.30 . 1 86 . 16 GLY N N 109.11 . 1 87 . 17 GLU H H 8.02 . 1 88 . 17 GLU HA H 4.29 . 1 89 . 17 GLU C C 175.62 . 1 90 . 17 GLU CA C 56.03 . 1 91 . 17 GLU CB C 30.41 . 1 92 . 17 GLU N N 123.16 . 1 93 . 18 THR H H 8.80 . 1 94 . 18 THR HA H 4.90 . 1 95 . 18 THR C C 177.02 . 1 96 . 18 THR CA C 63.17 . 1 97 . 18 THR CB C 69.27 . 1 98 . 18 THR N N 120.01 . 1 99 . 19 LEU H H 10.12 . 1 100 . 19 LEU HA H 4.81 . 1 101 . 19 LEU C C 175.69 . 1 102 . 19 LEU CA C 53.40 . 1 103 . 19 LEU CB C 42.79 . 1 104 . 19 LEU N N 132.44 . 1 105 . 20 THR H H 8.96 . 1 106 . 20 THR HA H 4.97 . 1 107 . 20 THR C C 174.23 . 1 108 . 20 THR CA C 63.97 . 1 109 . 20 THR CB C 70.16 . 1 110 . 20 THR N N 123.88 . 1 111 . 21 THR H H 8.87 . 1 112 . 21 THR HA H 4.71 . 1 113 . 21 THR C C 171.96 . 1 114 . 21 THR CA C 60.27 . 1 115 . 21 THR CB C 70.50 . 1 116 . 21 THR N N 123.66 . 1 117 . 22 LYS H H 8.05 . 1 118 . 22 LYS HA H 5.62 . 1 119 . 22 LYS C C 176.57 . 1 120 . 22 LYS CA C 53.95 . 1 121 . 22 LYS CB C 36.10 . 1 122 . 22 LYS N N 118.92 . 1 123 . 23 GLY H H 8.93 . 1 124 . 23 GLY HA2 H 4.73 . 2 125 . 23 GLY HA3 H 3.32 . 2 126 . 23 GLY C C 171.11 . 1 127 . 23 GLY CA C 43.99 . 1 128 . 23 GLY N N 108.10 . 1 129 . 24 LYS H H 8.85 . 1 130 . 24 LYS HA H 4.71 . 1 131 . 24 LYS C C 176.48 . 1 132 . 24 LYS CA C 55.08 . 1 133 . 24 LYS CB C 33.47 . 1 134 . 24 LYS N N 122.11 . 1 135 . 25 VAL H H 8.50 . 1 136 . 25 VAL HA H 3.22 . 1 137 . 25 VAL C C 177.60 . 1 138 . 25 VAL CA C 65.81 . 1 139 . 25 VAL CB C 31.67 . 1 140 . 25 VAL N N 123.28 . 1 141 . 26 GLY H H 8.91 . 1 142 . 26 GLY HA2 H 4.56 . 2 143 . 26 GLY HA3 H 3.58 . 2 144 . 26 GLY C C 174.54 . 1 145 . 26 GLY CA C 45.26 . 1 146 . 26 GLY N N 117.60 . 1 147 . 27 ASP H H 8.24 . 1 148 . 27 ASP HA H 4.74 . 1 149 . 27 ASP C C 176.56 . 1 150 . 27 ASP CA C 54.77 . 1 151 . 27 ASP CB C 41.48 . 1 152 . 27 ASP N N 123.94 . 1 153 . 28 SER H H 9.80 . 1 154 . 28 SER HA H 4.36 . 1 155 . 28 SER CA C 57.18 . 1 156 . 28 SER CB C 66.03 . 1 157 . 28 SER N N 119.15 . 1 158 . 29 LEU H H 8.31 . 1 159 . 29 LEU HA H 4.58 . 1 160 . 29 LEU CA C 58.04 . 1 161 . 29 LEU CB C 41.04 . 1 162 . 29 LEU N N 119.60 . 1 163 . 30 LEU C C 175.32 . 1 164 . 30 LEU CA C 55.71 . 1 165 . 31 ASP H H 7.74 . 1 166 . 31 ASP HA H 4.38 . 1 167 . 31 ASP C C 177.78 . 1 168 . 31 ASP CA C 58.05 . 1 169 . 31 ASP N N 121.84 . 1 170 . 32 VAL H H 7.82 . 1 171 . 32 VAL HA H 3.85 . 1 172 . 32 VAL C C 178.88 . 1 173 . 32 VAL CA C 66.27 . 1 174 . 32 VAL CB C 32.19 . 1 175 . 32 VAL N N 117.97 . 1 176 . 33 VAL H H 7.52 . 1 177 . 33 VAL HA H 3.06 . 1 178 . 33 VAL C C 178.01 . 1 179 . 33 VAL CA C 67.33 . 1 180 . 33 VAL CB C 32.31 . 1 181 . 33 VAL N N 122.43 . 1 182 . 34 VAL H H 8.48 . 1 183 . 34 VAL HA H 3.76 . 1 184 . 34 VAL C C 181.07 . 1 185 . 34 VAL CA C 66.36 . 1 186 . 34 VAL CB C 32.57 . 1 187 . 34 VAL N N 119.47 . 1 188 . 35 GLU H H 9.22 . 1 189 . 35 GLU HA H 4.03 . 1 190 . 35 GLU C C 177.91 . 1 191 . 35 GLU CA C 59.35 . 1 192 . 35 GLU CB C 29.41 . 1 193 . 35 GLU N N 121.19 . 1 194 . 36 ASN H H 7.21 . 1 195 . 36 ASN HA H 4.73 . 1 196 . 36 ASN C C 173.10 . 1 197 . 36 ASN CA C 53.51 . 1 198 . 36 ASN CB C 39.68 . 1 199 . 36 ASN N N 112.04 . 1 200 . 37 ASN H H 7.90 . 1 201 . 37 ASN HA H 4.40 . 1 202 . 37 ASN C C 174.88 . 1 203 . 37 ASN CA C 54.08 . 1 204 . 37 ASN CB C 37.43 . 1 205 . 37 ASN N N 118.37 . 1 206 . 38 LEU H H 7.84 . 1 207 . 38 LEU HA H 4.02 . 1 208 . 38 LEU C C 178.19 . 1 209 . 38 LEU CA C 55.57 . 1 210 . 38 LEU CB C 42.11 . 1 211 . 38 LEU N N 115.97 . 1 212 . 39 ASP H H 8.68 . 1 213 . 39 ASP HA H 4.55 . 1 214 . 39 ASP C C 174.70 . 1 215 . 39 ASP CA C 53.53 . 1 216 . 39 ASP CB C 39.41 . 1 217 . 39 ASP N N 123.55 . 1 218 . 40 ILE H H 7.67 . 1 219 . 40 ILE HA H 4.20 . 1 220 . 40 ILE C C 175.98 . 1 221 . 40 ILE CA C 60.09 . 1 222 . 40 ILE CB C 38.70 . 1 223 . 40 ILE N N 124.76 . 1 224 . 41 ASP H H 8.63 . 1 225 . 41 ASP HA H 4.37 . 1 226 . 41 ASP C C 178.43 . 1 227 . 41 ASP CA C 56.93 . 1 228 . 41 ASP CB C 41.64 . 1 229 . 41 ASP N N 128.35 . 1 230 . 42 GLY H H 9.15 . 1 231 . 42 GLY HA2 H 4.11 . 2 232 . 42 GLY HA3 H 3.80 . 2 233 . 42 GLY C C 174.08 . 1 234 . 42 GLY CA C 46.24 . 1 235 . 42 GLY N N 114.75 . 1 236 . 43 PHE H H 7.71 . 1 237 . 43 PHE HA H 4.60 . 1 238 . 43 PHE CA C 59.42 . 1 239 . 43 PHE CB C 41.05 . 1 240 . 43 PHE N N 121.68 . 1 241 . 59 PHE H H 8.79 . 1 242 . 59 PHE HA H 4.56 . 1 243 . 59 PHE C C 175.60 . 1 244 . 59 PHE CA C 58.59 . 1 245 . 59 PHE CB C 43.53 . 1 246 . 59 PHE N N 128.40 . 1 247 . 60 GLU H H 9.11 . 1 248 . 60 GLU HA H 4.36 . 1 249 . 60 GLU C C 178.88 . 1 250 . 60 GLU CA C 56.93 . 1 251 . 60 GLU CB C 32.33 . 1 252 . 60 GLU N N 119.55 . 1 253 . 61 ASP H H 9.41 . 1 254 . 61 ASP HA H 4.13 . 1 255 . 61 ASP C C 177.64 . 1 256 . 61 ASP CA C 59.14 . 1 257 . 61 ASP CB C 41.10 . 1 258 . 61 ASP N N 124.59 . 1 259 . 62 HIS H H 8.55 . 1 260 . 62 HIS HA H 4.52 . 1 261 . 62 HIS C C 176.96 . 1 262 . 62 HIS CA C 58.75 . 1 263 . 62 HIS CB C 29.58 . 1 264 . 62 HIS N N 114.63 . 1 265 . 63 ILE H H 6.33 . 1 266 . 63 ILE HA H 3.89 . 1 267 . 63 ILE C C 178.13 . 1 268 . 63 ILE CA C 59.92 . 1 269 . 63 ILE CB C 36.53 . 1 270 . 63 ILE N N 120.80 . 1 271 . 64 TYR H H 8.72 . 1 272 . 64 TYR HA H 3.58 . 1 273 . 64 TYR C C 177.70 . 1 274 . 64 TYR CA C 61.76 . 1 275 . 64 TYR CB C 39.66 . 1 276 . 64 TYR N N 120.57 . 1 277 . 65 GLU H H 7.88 . 1 278 . 65 GLU HA H 4.14 . 1 279 . 65 GLU C C 176.53 . 1 280 . 65 GLU CA C 58.18 . 1 281 . 65 GLU CB C 30.02 . 1 282 . 65 GLU N N 113.85 . 1 283 . 66 LYS H H 7.22 . 1 284 . 66 LYS HA H 4.48 . 1 285 . 66 LYS C C 176.84 . 1 286 . 66 LYS CA C 54.80 . 1 287 . 66 LYS CB C 33.34 . 1 288 . 66 LYS N N 116.52 . 1 289 . 67 LEU H H 6.85 . 1 290 . 67 LEU HA H 4.06 . 1 291 . 67 LEU C C 177.19 . 1 292 . 67 LEU CA C 54.69 . 1 293 . 67 LEU CB C 41.50 . 1 294 . 67 LEU N N 120.64 . 1 295 . 68 ASP H H 9.18 . 1 296 . 68 ASP HA H 4.32 . 1 297 . 68 ASP C C 175.60 . 1 298 . 68 ASP CA C 54.72 . 1 299 . 68 ASP CB C 40.96 . 1 300 . 68 ASP N N 123.38 . 1 301 . 69 ALA H H 8.30 . 1 302 . 69 ALA HA H 4.00 . 1 303 . 69 ALA C C 178.45 . 1 304 . 69 ALA CA C 52.68 . 1 305 . 69 ALA CB C 18.81 . 1 306 . 69 ALA N N 123.03 . 1 307 . 70 ILE H H 8.52 . 1 308 . 70 ILE HA H 4.67 . 1 309 . 70 ILE C C 176.75 . 1 310 . 70 ILE CA C 63.23 . 1 311 . 70 ILE CB C 39.35 . 1 312 . 70 ILE N N 125.25 . 1 313 . 71 THR H H 8.25 . 1 314 . 71 THR HA H 4.51 . 1 315 . 71 THR C C 175.68 . 1 316 . 71 THR CA C 61.18 . 1 317 . 71 THR CB C 72.33 . 1 318 . 71 THR N N 120.05 . 1 319 . 72 ASP H H 9.12 . 1 320 . 72 ASP HA H 4.31 . 1 321 . 72 ASP C C 177.82 . 1 322 . 72 ASP CA C 57.88 . 1 323 . 72 ASP CB C 40.20 . 1 324 . 72 ASP N N 123.46 . 1 325 . 73 GLU H H 9.00 . 1 326 . 73 GLU HA H 4.05 . 1 327 . 73 GLU C C 179.58 . 1 328 . 73 GLU CA C 60.74 . 1 329 . 73 GLU CB C 29.32 . 1 330 . 73 GLU N N 117.93 . 1 331 . 74 GLU H H 7.57 . 1 332 . 74 GLU HA H 4.02 . 1 333 . 74 GLU C C 178.11 . 1 334 . 74 GLU CA C 59.43 . 1 335 . 74 GLU CB C 29.17 . 1 336 . 74 GLU N N 120.30 . 1 337 . 75 ASN H H 8.65 . 1 338 . 75 ASN HA H 4.36 . 1 339 . 75 ASN C C 177.12 . 1 340 . 75 ASN CA C 57.20 . 1 341 . 75 ASN CB C 39.16 . 1 342 . 75 ASN N N 120.64 . 1 343 . 76 ASP H H 8.37 . 1 344 . 76 ASP HA H 4.32 . 1 345 . 76 ASP C C 178.76 . 1 346 . 76 ASP CA C 57.34 . 1 347 . 76 ASP CB C 40.42 . 1 348 . 76 ASP N N 117.86 . 1 349 . 77 MET H H 7.05 . 1 350 . 77 MET HA H 4.57 . 1 351 . 77 MET C C 180.05 . 1 352 . 77 MET CA C 55.78 . 1 353 . 77 MET CB C 34.85 . 1 354 . 77 MET N N 116.67 . 1 355 . 78 LEU H H 8.93 . 1 356 . 78 LEU HA H 3.83 . 1 357 . 78 LEU C C 178.64 . 1 358 . 78 LEU CA C 57.96 . 1 359 . 78 LEU CB C 41.61 . 1 360 . 78 LEU N N 124.74 . 1 361 . 79 ASP H H 8.04 . 1 362 . 79 ASP HA H 4.40 . 1 363 . 79 ASP C C 177.78 . 1 364 . 79 ASP CA C 56.91 . 1 365 . 79 ASP CB C 41.24 . 1 366 . 79 ASP N N 116.31 . 1 367 . 80 LEU H H 7.07 . 1 368 . 80 LEU HA H 4.39 . 1 369 . 80 LEU C C 177.44 . 1 370 . 80 LEU CA C 53.99 . 1 371 . 80 LEU CB C 42.60 . 1 372 . 80 LEU N N 117.42 . 1 373 . 81 ALA H H 7.92 . 1 374 . 81 ALA HA H 4.22 . 1 375 . 81 ALA C C 177.30 . 1 376 . 81 ALA CA C 52.51 . 1 377 . 81 ALA CB C 19.32 . 1 378 . 81 ALA N N 125.16 . 1 379 . 82 TYR H H 8.43 . 1 380 . 82 TYR HA H 4.46 . 1 381 . 82 TYR C C 177.02 . 1 382 . 82 TYR CA C 58.42 . 1 383 . 82 TYR CB C 38.96 . 1 384 . 82 TYR N N 123.92 . 1 385 . 83 GLY H H 8.52 . 1 386 . 83 GLY HA2 H 4.97 . 2 387 . 83 GLY HA3 H 3.71 . 2 388 . 83 GLY C C 174.76 . 1 389 . 83 GLY CA C 46.55 . 1 390 . 83 GLY N N 113.66 . 1 391 . 84 LEU H H 7.31 . 1 392 . 84 LEU HA H 3.89 . 1 393 . 84 LEU C C 176.69 . 1 394 . 84 LEU CA C 57.00 . 1 395 . 84 LEU CB C 42.79 . 1 396 . 84 LEU N N 119.14 . 1 397 . 85 THR H H 8.93 . 1 398 . 85 THR HA H 4.97 . 1 399 . 85 THR C C 174.88 . 1 400 . 85 THR CA C 60.16 . 1 401 . 85 THR CB C 74.50 . 1 402 . 85 THR N N 118.66 . 1 403 . 86 ASP H H 8.57 . 1 404 . 86 ASP HA H 4.55 . 1 405 . 86 ASP C C 176.09 . 1 406 . 86 ASP CA C 54.29 . 1 407 . 86 ASP CB C 39.74 . 1 408 . 86 ASP N N 118.69 . 1 409 . 87 ARG H H 8.39 . 1 410 . 87 ARG HA H 4.26 . 1 411 . 87 ARG C C 175.75 . 1 412 . 87 ARG CA C 53.73 . 1 413 . 87 ARG CB C 30.02 . 1 414 . 87 ARG N N 120.57 . 1 415 . 88 SER H H 7.07 . 1 416 . 88 SER HA H 5.48 . 1 417 . 88 SER C C 174.64 . 1 418 . 88 SER CA C 60.72 . 1 419 . 88 SER CB C 65.48 . 1 420 . 88 SER N N 120.81 . 1 421 . 89 ARG H H 9.46 . 1 422 . 89 ARG HA H 4.87 . 1 423 . 89 ARG CA C 52.93 . 1 424 . 89 ARG CB C 34.17 . 1 425 . 89 ARG N N 118.98 . 1 426 . 94 ILE HA H 4.25 . 1 427 . 94 ILE C C 174.06 . 1 428 . 94 ILE CA C 58.01 . 1 429 . 94 ILE CB C 37.05 . 1 430 . 95 CYS H H 8.20 . 1 431 . 95 CYS HA H 5.00 . 1 432 . 95 CYS C C 175.16 . 1 433 . 95 CYS CA C 57.50 . 1 434 . 95 CYS CB C 29.33 . 1 435 . 95 CYS N N 124.34 . 1 436 . 96 LEU H H 9.05 . 1 437 . 96 LEU HA H 4.76 . 1 438 . 96 LEU C C 178.97 . 1 439 . 96 LEU CA C 55.64 . 1 440 . 96 LEU CB C 42.46 . 1 441 . 96 LEU N N 121.35 . 1 442 . 97 THR H H 7.86 . 1 443 . 97 THR HA H 4.68 . 1 444 . 97 THR C C 175.30 . 1 445 . 97 THR CA C 59.42 . 1 446 . 97 THR CB C 71.69 . 1 447 . 97 THR N N 114.58 . 1 448 . 98 LYS H H 9.28 . 1 449 . 98 LYS HA H 3.92 . 1 450 . 98 LYS C C 179.19 . 1 451 . 98 LYS CA C 59.55 . 1 452 . 98 LYS CB C 32.08 . 1 453 . 98 LYS N N 122.12 . 1 454 . 99 SER H H 8.22 . 1 455 . 99 SER HA H 4.30 . 1 456 . 99 SER C C 174.77 . 1 457 . 99 SER CA C 60.42 . 1 458 . 99 SER CB C 62.69 . 1 459 . 99 SER N N 113.39 . 1 460 . 100 MET H H 7.73 . 1 461 . 100 MET HA H 4.16 . 1 462 . 100 MET C C 174.12 . 1 463 . 100 MET CA C 56.92 . 1 464 . 100 MET CB C 32.37 . 1 465 . 100 MET N N 121.20 . 1 466 . 101 ASP H H 7.27 . 1 467 . 101 ASP HA H 4.41 . 1 468 . 101 ASP C C 177.82 . 1 469 . 101 ASP CA C 57.02 . 1 470 . 101 ASP CB C 40.94 . 1 471 . 101 ASP N N 118.59 . 1 472 . 102 ASN H H 10.06 . 1 473 . 102 ASN HA H 3.88 . 1 474 . 102 ASN C C 174.34 . 1 475 . 102 ASN CA C 55.90 . 1 476 . 102 ASN CB C 38.01 . 1 477 . 102 ASN N N 123.28 . 1 478 . 103 MET H H 8.48 . 1 479 . 103 MET HA H 4.70 . 1 480 . 103 MET C C 173.08 . 1 481 . 103 MET CA C 56.67 . 1 482 . 103 MET CB C 31.45 . 1 483 . 103 MET N N 118.84 . 1 484 . 104 THR H H 8.69 . 1 485 . 104 THR HA H 5.47 . 1 486 . 104 THR C C 175.97 . 1 487 . 104 THR CA C 61.83 . 1 488 . 104 THR CB C 70.59 . 1 489 . 104 THR N N 117.57 . 1 490 . 105 VAL H H 9.19 . 1 491 . 105 VAL HA H 4.62 . 1 492 . 105 VAL CA C 58.32 . 1 493 . 105 VAL CB C 34.48 . 1 494 . 105 VAL N N 123.32 . 1 495 . 106 ARG H H 8.44 . 1 496 . 106 ARG HA H 4.31 . 1 497 . 106 ARG CA C 54.57 . 1 498 . 106 ARG CB C 33.96 . 1 499 . 106 ARG N N 121.33 . 1 500 . 108 PRO HA H 4.30 . 1 501 . 108 PRO C C 176.32 . 1 502 . 108 PRO CA C 62.95 . 1 503 . 108 PRO CB C 32.38 . 1 504 . 109 GLU H H 8.69 . 1 505 . 109 GLU HA H 4.22 . 1 506 . 109 GLU C C 176.48 . 1 507 . 109 GLU CA C 57.22 . 1 508 . 109 GLU CB C 30.91 . 1 509 . 109 GLU N N 121.77 . 1 510 . 110 THR H H 8.38 . 1 511 . 110 THR HA H 4.35 . 1 512 . 110 THR C C 174.09 . 1 513 . 110 THR CA C 61.51 . 1 514 . 110 THR CB C 70.24 . 1 515 . 110 THR N N 115.08 . 1 516 . 111 VAL H H 7.95 . 1 517 . 111 VAL HA H 3.93 . 1 518 . 111 VAL C C 175.46 . 1 519 . 111 VAL CA C 61.87 . 1 520 . 111 VAL CB C 33.20 . 1 521 . 111 VAL N N 122.76 . 1 522 . 112 ALA H H 8.34 . 1 523 . 112 ALA HA H 4.24 . 1 524 . 112 ALA C C 177.55 . 1 525 . 112 ALA CA C 52.47 . 1 526 . 112 ALA CB C 19.67 . 1 527 . 112 ALA N N 129.12 . 1 528 . 113 ASP H H 8.39 . 1 529 . 113 ASP HA H 4.53 . 1 530 . 113 ASP C C 175.09 . 1 531 . 113 ASP CA C 54.62 . 1 532 . 113 ASP CB C 41.16 . 1 533 . 113 ASP N N 121.32 . 1 534 . 114 ALA H H 7.91 . 1 535 . 114 ALA HA H 4.09 . 1 536 . 114 ALA CA C 54.03 . 1 537 . 114 ALA CB C 20.38 . 1 538 . 114 ALA N N 130.28 . 1 stop_ save_