data_25000 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; WW3 domain of Nedd4L in complex with its HECT domain PY motif ; _BMRB_accession_number 25000 _BMRB_flat_file_name bmr25000.str _Entry_type original _Submission_date 2014-06-02 _Accession_date 2014-06-02 _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 Escobedo Albert . . 2 Macias Maria J. . 3 Gomes Tiago . . 4 Aragon Eric . . 5 Martin-Malpartida Pau . . 6 Ruiz Lidia . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 3 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 246 "13C chemical shifts" 59 "15N chemical shifts" 27 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2014-10-20 original author . stop_ _Original_release_date 2014-10-20 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'Structural Basis of the Activation and Degradation Mechanisms of the E3 Ubiquitin Ligase Nedd4L' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 25295397 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Escobedo Albert . . 2 Gomes Tiago . . 3 Aragon Eric . . 4 Martin-Malpartida Pau . . 5 Ruiz Lidia . . 6 Macias Maria . . stop_ _Journal_abbreviation Structure _Journal_volume 22 _Journal_issue 10 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 1446 _Page_last 1457 _Year 2014 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'WW3 domain of Nedd4L in complex with its HECT domain PY motif' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label WW3 $WW3 HECT_PY $HECT_PY stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_WW3 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common WW3 _Molecular_mass 3833.352 _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 48 _Mol_residue_sequence ; GAMEQSFLPPGWEMRIAPNG RPFFIDHNTKTTTWEDPRLK FPVHMRSK ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 472 GLY 2 473 ALA 3 474 MET 4 475 GLU 5 476 GLN 6 477 SER 7 478 PHE 8 479 LEU 9 480 PRO 10 481 PRO 11 482 GLY 12 483 TRP 13 484 GLU 14 485 MET 15 486 ARG 16 487 ILE 17 488 ALA 18 489 PRO 19 490 ASN 20 491 GLY 21 492 ARG 22 493 PRO 23 494 PHE 24 495 PHE 25 496 ILE 26 497 ASP 27 498 HIS 28 499 ASN 29 500 THR 30 501 LYS 31 502 THR 32 503 THR 33 504 THR 34 505 TRP 35 506 GLU 36 507 ASP 37 508 PRO 38 509 ARG 39 510 LEU 40 511 LYS 41 512 PHE 42 513 PRO 43 514 VAL 44 515 HIS 45 516 MET 46 517 ARG 47 518 SER 48 519 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-01-29 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 17529 NEDD4LWW3 91.67 44 97.73 97.73 2.95e-23 PDB 1WR7 "Solution Structure Of The Third Ww Domain Of Nedd4-2" 75.00 41 100.00 100.00 1.73e-17 PDB 2LAJ "Third Ww Domain Of Human Nedd4l In Complex With Doubly Phosphorylated Human Smad3 Derived Peptide" 91.67 44 97.73 97.73 2.95e-23 PDB 2MPT "Ww3 Domain Of Nedd4l In Complex With Its Hect Domain Py Motif" 100.00 48 100.00 100.00 6.45e-27 stop_ save_ save_HECT_PY _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common HECT_PY _Molecular_mass 1608.784 _Mol_thiol_state 'not present' _Details . _Residue_count 16 _Mol_residue_sequence RLDLPPYETFEDLREK loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 925 ARG 2 926 LEU 3 927 ASP 4 928 LEU 5 929 PRO 6 930 PRO 7 931 TYR 8 932 GLU 9 933 THR 10 934 PHE 11 935 GLU 12 936 ASP 13 937 LEU 14 938 ARG 15 939 GLU 16 940 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-01-29 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 2ONI "Catalytic Domain Of The Human Nedd4-Like E3 Ligase" 100.00 392 100.00 100.00 9.18e-01 PDB 3JVZ E2~ubiquitin-Hect 100.00 385 100.00 100.00 8.03e-01 PDB 3JW0 E2~ubiquitin-Hect 100.00 385 100.00 100.00 8.03e-01 DBJ BAA23711 "KIAA0439 [Homo sapiens]" 100.00 995 100.00 100.00 1.27e+00 DBJ BAB69424 "NEDD4-like ubiquitin ligase 3 [Homo sapiens]" 100.00 955 100.00 100.00 1.19e+00 DBJ BAC31307 "unnamed protein product [Mus musculus]" 100.00 835 100.00 100.00 1.13e+00 DBJ BAE37373 "unnamed protein product [Mus musculus]" 100.00 896 100.00 100.00 1.23e+00 DBJ BAG11191 "E3 ubiquitin-protein ligase NEDD4-like protein [synthetic construct]" 100.00 834 100.00 100.00 1.17e+00 EMBL CAB70754 "hypothetical protein [Homo sapiens]" 100.00 820 100.00 100.00 1.29e+00 EMBL CAH90908 "hypothetical protein [Pongo abelii]" 100.00 959 100.00 100.00 1.16e+00 GB AAG43524 "NEDD4La [Homo sapiens]" 100.00 854 100.00 100.00 1.11e+00 GB AAH00621 "NEDD4L protein, partial [Homo sapiens]" 100.00 855 100.00 100.00 1.22e+00 GB AAH19345 "Unknown (protein for IMAGE:3604024), partial [Homo sapiens]" 100.00 855 100.00 100.00 1.22e+00 GB AAH32597 "NEDD4L protein [Homo sapiens]" 100.00 911 100.00 100.00 1.24e+00 GB AAH39746 "Neural precursor cell expressed, developmentally down-regulated gene 4-like [Mus musculus]" 100.00 855 100.00 100.00 1.09e+00 REF NP_001008301 "E3 ubiquitin-protein ligase NEDD4-like [Rattus norvegicus]" 100.00 963 100.00 100.00 1.11e+00 REF NP_001107858 "E3 ubiquitin-protein ligase NEDD4-like isoform 1 [Mus musculus]" 100.00 976 100.00 100.00 1.13e+00 REF NP_001125518 "E3 ubiquitin-protein ligase NEDD4-like [Pongo abelii]" 100.00 959 100.00 100.00 1.16e+00 REF NP_001138436 "E3 ubiquitin-protein ligase NEDD4-like isoform 2 [Homo sapiens]" 100.00 854 100.00 100.00 1.11e+00 REF NP_001138437 "E3 ubiquitin-protein ligase NEDD4-like isoform 2 [Homo sapiens]" 100.00 854 100.00 100.00 1.11e+00 SP Q5RBF2 "RecName: Full=E3 ubiquitin-protein ligase NEDD4-like [Pongo abelii]" 100.00 959 100.00 100.00 1.16e+00 SP Q8CFI0 "RecName: Full=E3 ubiquitin-protein ligase NEDD4-like; AltName: Full=NEDD4.2; AltName: Full=Nedd4-2 [Mus musculus]" 100.00 1004 100.00 100.00 1.09e+00 SP Q96PU5 "RecName: Full=E3 ubiquitin-protein ligase NEDD4-like; AltName: Full=NEDD4.2; AltName: Full=Nedd4-2 [Homo sapiens]" 100.00 975 100.00 100.00 1.15e+00 TPG DAA15876 "TPA: neural precursor cell expressed, developmentally down-regulated 4-like [Bos taurus]" 100.00 854 100.00 100.00 1.10e+00 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $WW3 Human 9606 Eukaryota Metazoa Homo sapiens $HECT_PY Human 9606 Eukaryota Metazoa 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_name $WW3 'recombinant technology' . . . BL21 DE3 pETM11 $HECT_PY 'chemical synthesis' . . . . . . 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 $WW3 0.7 mM 'natural abundance' $HECT_PY 1.4 mM 'natural abundance' 'sodium chloride' 150 mM 'natural abundance' 'sodium phosphate' 20 mM 'natural abundance' H2O 90 % 'natural abundance' D2O 10 % 'natural abundance' stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $WW3 1 mM '[U-100% 15N]' $HECT_PY 2 mM 'natural abundance' 'sodium chloride' 150 mM 'natural abundance' 'sodium phosphate' 20 mM 'natural abundance' H2O 90 % 'natural abundance' D2O 10 % 'natural abundance' stop_ save_ save_sample_3 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $WW3 0.8 mM '[U-100% 13C; U-100% 15N]' $HECT_PY 1.6 mM 'natural abundance' 'sodium chloride' 150 mM 'natural abundance' 'sodium phosphate' 20 mM 'natural abundance' H2O 90 % 'natural abundance' D2O 10 % 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_CNSSOLVE _Saveframe_category software _Name CNSSOLVE _Version . loop_ _Vendor _Address _Electronic_address 'Brunger, Adams, Clore, Gros, Nilges and Read' . . stop_ loop_ _Task refinement 'structure solution' stop_ _Details . save_ save_CARA _Saveframe_category software _Name CARA _Version . loop_ _Vendor _Address _Electronic_address 'Keller and Wuthrich' . . stop_ loop_ _Task 'chemical shift assignment' 'peak picking' stop_ _Details . save_ save_NMRPipe _Saveframe_category software _Name NMRPipe _Version . loop_ _Vendor _Address _Electronic_address 'Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax' . . stop_ loop_ _Task processing stop_ _Details . save_ save_TOPSPIN _Saveframe_category software _Name TOPSPIN _Version . loop_ _Vendor _Address _Electronic_address 'Bruker Biospin' . . stop_ loop_ _Task collection stop_ _Details . save_ save_TALOS _Saveframe_category software _Name TALOS _Version . loop_ _Vendor _Address _Electronic_address 'Cornilescu, Delaglio and Bax' . . stop_ loop_ _Task 'geometry optimization' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-1H_NOESY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $sample_1 save_ save_2D_1H-1H_TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label $sample_1 save_ save_3D_1H-15N_NOESY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-15N NOESY' _Sample_label $sample_2 save_ save_3D_1H-15N_TOCSY_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-15N TOCSY' _Sample_label $sample_2 save_ save_3D_HNCACB_5 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCACB' _Sample_label $sample_3 save_ save_3D_CBCA(CO)NH_6 _Saveframe_category NMR_applied_experiment _Experiment_name '3D CBCA(CO)NH' _Sample_label $sample_3 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units temperature 298 . K pH 7 . pH pressure 1 . atm stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference_1 _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 0.00 internal direct . . . 1.000000000 DSS C 13 'methyl protons' ppm 0.00 na indirect . . . 0.251449530 DSS N 15 'methyl protons' ppm 0.00 na indirect . . . 0.101329118 stop_ save_ ################################### # Assigned chemical shift lists # ################################### ################################################################### # Chemical Shift Ambiguity Index Value Definitions # # # # The values other than 1 are used for those atoms with different # # chemical shifts that cannot be assigned to stereospecific atoms # # or to specific residues or chains. # # # # Index Value Definition # # # # 1 Unique (including isolated methyl protons, # # geminal atoms, and geminal methyl # # groups with identical chemical shifts) # # (e.g. ILE HD11, HD12, HD13 protons) # # 2 Ambiguity of geminal atoms or geminal methyl # # proton groups (e.g. ASP HB2 and HB3 # # protons, LEU CD1 and CD2 carbons, or # # LEU HD11, HD12, HD13 and HD21, HD22, # # HD23 methyl protons) # # 3 Aromatic atoms on opposite sides of # # symmetrical rings (e.g. TYR HE1 and HE2 # # protons) # # 4 Intraresidue ambiguities (e.g. LYS HG and # # HD protons or TRP HZ2 and HZ3 protons) # # 5 Interresidue ambiguities (LYS 12 vs. LYS 27) # # 6 Intermolecular ambiguities (e.g. ASP 31 CA # # in monomer 1 and ASP 31 CA in monomer 2 # # of an asymmetrical homodimer, duplex # # DNA assignments, or other assignments # # that may apply to atoms in one or more # # molecule in the molecular assembly) # # 9 Ambiguous, specific ambiguity not defined # # # ################################################################### save_assigned_chem_shift_list_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-1H NOESY' '2D 1H-1H TOCSY' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name WW3 _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 1 478 7 PHE H H 7.971 0.1 1 2 478 7 PHE HA H 4.567 0.1 1 3 478 7 PHE HB2 H 3.135 0.1 2 4 478 7 PHE HB3 H 2.901 0.1 2 5 478 7 PHE HD1 H 7.174 0.1 3 6 478 7 PHE HE1 H 7.244 0.1 3 7 479 8 LEU H H 8.384 0.1 1 8 479 8 LEU HA H 4.190 0.1 1 9 479 8 LEU HB2 H 1.706 0.002 2 10 479 8 LEU HB3 H 1.615 0.1 2 11 479 8 LEU HG H 1.339 0.1 1 12 479 8 LEU HD1 H 0.978 0.1 2 13 479 8 LEU HD2 H 0.748 0.1 2 14 480 9 PRO HA H 4.570 0.001 1 15 480 9 PRO HB2 H 2.291 0.1 2 16 480 9 PRO HB3 H 2.068 0.034 2 17 480 9 PRO HG2 H 1.198 0.1 2 18 480 9 PRO HD2 H 3.754 0.001 2 19 480 9 PRO HD3 H 3.497 0.1 2 20 482 11 GLY H H 8.995 0.1 1 21 482 11 GLY HA2 H 3.787 0.1 2 22 482 11 GLY HA3 H 4.185 0.1 2 23 483 12 TRP H H 7.709 0.001 1 24 483 12 TRP HA H 5.987 0.1 1 25 483 12 TRP HB2 H 3.247 0.1 2 26 483 12 TRP HB3 H 3.020 0.1 2 27 483 12 TRP HD1 H 6.972 0.1 1 28 483 12 TRP HE1 H 10.240 0.1 1 29 483 12 TRP HE3 H 7.325 0.1 1 30 483 12 TRP HZ2 H 7.283 0.1 1 31 483 12 TRP HZ3 H 6.779 0.001 1 32 483 12 TRP HH2 H 7.063 0.001 1 33 484 13 GLU H H 9.350 0.1 1 34 484 13 GLU HA H 4.712 0.1 1 35 484 13 GLU HB2 H 1.994 0.1 2 36 484 13 GLU HB3 H 1.915 0.001 2 37 484 13 GLU HG2 H 2.178 0.1 2 38 484 13 GLU HG3 H 2.112 0.001 2 39 485 14 MET H H 8.778 0.1 1 40 485 14 MET HA H 4.863 0.1 1 41 485 14 MET HB2 H 1.806 0.1 2 42 485 14 MET HG2 H 1.892 0.1 2 43 485 14 MET HE H 1.707 0.001 1 44 486 15 ARG H H 7.984 0.1 1 45 486 15 ARG HA H 4.282 0.1 1 46 486 15 ARG HB2 H 1.355 0.1 2 47 486 15 ARG HB3 H 0.393 0.1 2 48 486 15 ARG HG2 H 1.210 0.1 2 49 486 15 ARG HG3 H 0.987 0.1 2 50 486 15 ARG HD2 H 3.121 0.002 2 51 486 15 ARG HD3 H 2.717 0.001 2 52 486 15 ARG HH22 H 6.717 0.002 2 53 487 16 ILE H H 8.146 0.1 1 54 487 16 ILE HA H 4.347 0.001 1 55 487 16 ILE HB H 1.798 0.1 1 56 487 16 ILE HG12 H 1.203 0.1 2 57 487 16 ILE HG2 H 0.600 0.1 1 58 487 16 ILE HD1 H 0.863 0.1 1 59 488 17 ALA H H 9.043 0.1 1 60 488 17 ALA HA H 4.573 0.1 1 61 488 17 ALA HB H 1.850 0.001 1 62 489 18 PRO HA H 4.308 0.1 1 63 489 18 PRO HB2 H 2.372 0.002 2 64 489 18 PRO HD2 H 3.772 0.1 2 65 490 19 ASN H H 7.622 0.1 1 66 490 19 ASN HA H 4.498 0.1 1 67 490 19 ASN HB2 H 3.159 0.1 2 68 490 19 ASN HB3 H 2.789 0.001 2 69 490 19 ASN HD21 H 7.353 0.1 2 70 490 19 ASN HD22 H 5.603 0.003 2 71 491 20 GLY H H 8.600 0.1 1 72 491 20 GLY HA2 H 3.470 0.1 2 73 491 20 GLY HA3 H 4.214 0.1 2 74 492 21 ARG H H 7.879 0.1 1 75 492 21 ARG HA H 4.719 0.1 1 76 492 21 ARG HB2 H 1.916 0.1 2 77 492 21 ARG HB3 H 1.847 0.1 2 78 492 21 ARG HG2 H 1.631 0.001 2 79 492 21 ARG HE H 6.984 0.1 1 80 493 22 PRO HA H 5.111 0.002 1 81 493 22 PRO HB2 H 2.224 0.1 2 82 493 22 PRO HB3 H 2.021 0.1 2 83 493 22 PRO HG2 H 1.648 0.1 2 84 493 22 PRO HD2 H 4.013 0.1 2 85 493 22 PRO HD3 H 3.742 0.004 2 86 494 23 PHE H H 8.999 0.001 1 87 494 23 PHE HA H 4.908 0.1 1 88 494 23 PHE HB2 H 2.939 0.1 2 89 494 23 PHE HB3 H 2.377 0.1 2 90 494 23 PHE HD2 H 6.833 0.1 3 91 494 23 PHE HE2 H 6.715 0.1 3 92 495 24 PHE H H 8.999 0.1 1 93 495 24 PHE HA H 5.128 0.1 1 94 495 24 PHE HB2 H 3.230 0.1 2 95 495 24 PHE HB3 H 3.152 0.1 2 96 495 24 PHE HD2 H 7.001 0.1 3 97 495 24 PHE HE2 H 7.196 0.1 3 98 496 25 ILE H H 9.147 0.001 1 99 496 25 ILE HA H 4.042 0.1 1 100 496 25 ILE HB H 1.195 0.1 1 101 496 25 ILE HG12 H 0.950 0.1 2 102 496 25 ILE HG13 H 0.405 0.003 2 103 496 25 ILE HG2 H 0.052 0.1 1 104 496 25 ILE HD1 H 0.015 0.1 1 105 497 26 ASP H H 8.224 0.001 1 106 497 26 ASP HA H 3.635 0.1 1 107 497 26 ASP HB2 H 1.905 0.1 2 108 497 26 ASP HB3 H -0.039 0.1 2 109 498 27 HIS H H 8.754 0.001 1 110 498 27 HIS HA H 4.187 0.1 1 111 498 27 HIS HB2 H 3.230 0.1 2 112 498 27 HIS HB3 H 3.013 0.1 2 113 498 27 HIS HD2 H 6.812 0.1 1 114 498 27 HIS HE1 H 7.594 0.002 1 115 499 28 ASN H H 8.273 0.1 1 116 499 28 ASN HA H 4.297 0.001 1 117 499 28 ASN HB2 H 3.213 0.001 2 118 499 28 ASN HB3 H 2.565 0.002 2 119 499 28 ASN HD21 H 7.870 0.001 2 120 499 28 ASN HD22 H 7.722 0.1 2 121 500 29 THR H H 6.397 0.001 1 122 500 29 THR HA H 4.125 0.1 1 123 500 29 THR HG2 H 1.024 0.001 1 124 501 30 LYS H H 7.512 0.1 1 125 501 30 LYS HA H 2.121 0.1 1 126 501 30 LYS HB2 H 1.443 0.1 2 127 501 30 LYS HG2 H 0.886 0.1 2 128 501 30 LYS HG3 H 0.798 0.1 2 129 501 30 LYS HD2 H 0.967 0.1 2 130 501 30 LYS HE2 H 1.829 0.1 2 131 502 31 THR H H 7.137 0.001 1 132 502 31 THR HA H 4.685 0.1 1 133 502 31 THR HB H 3.921 0.1 1 134 502 31 THR HG2 H 0.981 0.1 1 135 503 32 THR H H 7.884 0.1 1 136 503 32 THR HA H 5.379 0.1 1 137 503 32 THR HB H 4.092 0.1 1 138 503 32 THR HG2 H 1.084 0.001 1 139 504 33 THR H H 9.122 0.001 1 140 504 33 THR HA H 4.693 0.1 1 141 504 33 THR HB H 4.397 0.1 1 142 504 33 THR HG2 H 1.575 0.1 1 143 505 34 TRP H H 8.742 0.002 1 144 505 34 TRP HA H 5.243 0.1 1 145 505 34 TRP HB2 H 3.648 0.1 2 146 505 34 TRP HB3 H 3.129 0.1 2 147 505 34 TRP HD1 H 7.278 0.001 1 148 505 34 TRP HE1 H 10.041 0.1 1 149 505 34 TRP HE3 H 8.264 0.1 1 150 505 34 TRP HZ2 H 7.084 0.1 1 151 505 34 TRP HZ3 H 6.978 0.1 1 152 505 34 TRP HH2 H 6.906 0.001 1 153 506 35 GLU H H 8.752 0.1 1 154 506 35 GLU HA H 4.141 0.1 1 155 506 35 GLU HB2 H 2.066 0.1 2 156 506 35 GLU HG2 H 2.211 0.1 2 157 507 36 ASP H H 8.022 0.002 1 158 507 36 ASP HA H 2.521 0.1 1 159 507 36 ASP HB2 H 2.184 0.1 2 160 508 37 PRO HA H 3.832 0.1 1 161 508 37 PRO HB2 H 0.766 0.1 2 162 508 37 PRO HB3 H 0.693 0.1 2 163 508 37 PRO HG2 H 0.537 0.1 2 164 508 37 PRO HG3 H 0.163 0.1 2 165 508 37 PRO HD2 H 2.684 0.001 2 166 509 38 ARG H H 8.193 0.1 1 167 509 38 ARG HA H 4.160 0.1 1 168 509 38 ARG HB2 H 1.758 0.1 2 169 509 38 ARG HB3 H 1.677 0.001 2 170 509 38 ARG HG2 H 1.544 0.003 2 171 509 38 ARG HD2 H 3.101 0.1 2 stop_ save_ save_assigned_chem_shift_list_2 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-1H NOESY' '2D 1H-1H TOCSY' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name HECT_PY _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 1 925 1 ARG H H 8.209 0.1 1 2 926 2 LEU H H 8.094 0.1 1 3 926 2 LEU HA H 3.983 0.1 1 4 926 2 LEU HB2 H 1.931 0.1 2 5 926 2 LEU HD1 H 0.808 0.1 2 6 927 3 ASP H H 7.845 0.1 1 7 927 3 ASP HA H 4.382 0.1 1 8 927 3 ASP HB2 H 3.069 0.1 2 9 927 3 ASP HB3 H 2.948 0.1 2 10 928 4 LEU HA H 3.164 0.1 1 11 928 4 LEU HB2 H 1.297 0.1 2 12 928 4 LEU HB3 H 0.949 0.1 2 13 928 4 LEU HD1 H 0.200 0.1 2 14 929 5 PRO HG2 H 1.354 0.1 2 15 929 5 PRO HD2 H 3.785 0.1 2 16 929 5 PRO HD3 H 3.501 0.1 2 17 931 7 TYR H H 8.433 0.1 1 18 931 7 TYR HA H 3.687 0.1 1 19 931 7 TYR HB2 H 2.655 0.1 2 20 931 7 TYR HB3 H 2.560 0.1 2 21 931 7 TYR HD1 H 6.641 0.1 3 22 931 7 TYR HE1 H 6.392 0.1 3 23 933 9 THR H H 7.518 0.1 1 24 933 9 THR HA H 3.953 0.1 1 25 933 9 THR HB H 3.677 0.1 1 26 933 9 THR HG2 H 0.881 0.1 1 27 934 10 PHE H H 7.887 0.1 1 28 934 10 PHE HA H 4.377 0.1 1 29 934 10 PHE HB2 H 3.069 0.1 2 30 934 10 PHE HB3 H 2.568 0.1 2 31 934 10 PHE HD2 H 7.024 0.1 3 32 934 10 PHE HE2 H 7.091 0.1 3 33 934 10 PHE HZ H 7.152 0.1 1 34 935 11 GLU H H 7.647 0.1 1 35 935 11 GLU HA H 4.014 0.1 1 36 935 11 GLU HB2 H 1.838 0.1 2 37 935 11 GLU HB3 H 1.780 0.1 2 38 935 11 GLU HG2 H 2.058 0.1 2 39 936 12 ASP H H 8.227 0.1 1 40 936 12 ASP HA H 4.440 0.1 1 41 936 12 ASP HB2 H 2.636 0.1 2 42 936 12 ASP HB3 H 2.508 0.1 2 43 937 13 LEU H H 8.062 0.1 1 44 937 13 LEU HA H 4.178 0.1 1 45 937 13 LEU HB2 H 1.530 0.1 2 46 937 13 LEU HG H 1.587 0.1 1 47 937 13 LEU HD1 H 0.819 0.1 2 48 937 13 LEU HD2 H 0.743 0.1 2 stop_ save_ save_assigned_chem_shift_list_3 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '3D HNCACB' '3D CBCA(CO)NH' stop_ loop_ _Sample_label $sample_3 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name WW3 _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 1 478 7 PHE H H 8.062 0.050 1 2 478 7 PHE CA C 54.686 0.070 1 3 478 7 PHE CB C 36.756 0.035 1 4 478 7 PHE N N 121.137 0.050 1 5 479 8 LEU H H 8.404 0.050 1 6 479 8 LEU CA C 50.795 0.050 1 7 479 8 LEU CB C 39.196 0.050 1 8 479 8 LEU N N 122.454 0.050 1 9 481 10 PRO CA C 62.041 0.050 1 10 481 10 PRO CB C 29.224 0.050 1 11 482 11 GLY H H 9.021 0.050 1 12 482 11 GLY CA C 43.404 0.035 1 13 482 11 GLY N N 113.164 0.050 1 14 483 12 TRP H H 7.824 0.050 1 15 483 12 TRP CA C 54.190 0.071 1 16 483 12 TRP CB C 29.436 0.050 1 17 483 12 TRP N N 118.566 0.050 1 18 484 13 GLU H H 9.400 0.050 1 19 484 13 GLU CA C 52.634 0.071 1 20 484 13 GLU CB C 33.184 0.050 1 21 484 13 GLU N N 120.909 0.050 1 22 485 14 MET H H 8.893 0.050 1 23 485 14 MET CA C 51.750 0.248 1 24 485 14 MET CB C 33.149 0.530 1 25 485 14 MET N N 125.140 0.050 1 26 486 15 ARG H H 8.150 0.050 1 27 486 15 ARG CA C 51.574 0.142 1 28 486 15 ARG CB C 31.982 0.050 1 29 486 15 ARG N N 126.181 0.050 1 30 487 16 ILE H H 8.191 0.050 1 31 487 16 ILE CA C 57.232 0.050 1 32 487 16 ILE CB C 34.988 0.177 1 33 487 16 ILE N N 119.543 0.050 1 34 488 17 ALA H H 9.028 0.050 1 35 488 17 ALA CA C 49.452 0.050 1 36 488 17 ALA CB C 15.361 0.050 1 37 488 17 ALA N N 131.616 0.050 1 38 489 18 PRO CA C 63.244 0.050 1 39 489 18 PRO CB C 29.294 0.050 1 40 490 19 ASN H H 7.602 0.050 1 41 490 19 ASN CA C 50.017 0.050 1 42 490 19 ASN CB C 34.847 0.035 1 43 490 19 ASN N N 112.091 0.050 1 44 491 20 GLY H H 8.669 0.050 1 45 491 20 GLY CA C 42.626 0.035 1 46 491 20 GLY N N 109.097 0.050 1 47 492 21 ARG H H 7.824 0.050 1 48 492 21 ARG CA C 51.361 0.050 1 49 492 21 ARG CB C 28.870 0.050 1 50 492 21 ARG N N 123.252 0.050 1 51 493 22 PRO CA C 60.485 0.050 1 52 493 22 PRO CB C 30.143 0.050 1 53 494 23 PHE H H 8.967 0.050 1 54 494 23 PHE CA C 53.341 0.070 1 55 494 23 PHE CB C 39.055 0.424 1 56 494 23 PHE N N 118.469 0.050 1 57 495 24 PHE H H 9.022 0.050 1 58 495 24 PHE CA C 54.757 0.141 1 59 495 24 PHE CB C 38.453 0.107 1 60 495 24 PHE N N 117.362 0.050 1 61 496 25 ILE H H 9.234 0.050 1 62 496 25 ILE CA C 58.293 0.050 1 63 496 25 ILE CB C 39.054 0.142 1 64 496 25 ILE N N 123.480 0.050 1 65 497 26 ASP H H 8.261 0.050 1 66 497 26 ASP CA C 48.922 0.035 1 67 497 26 ASP CB C 36.897 0.035 1 68 497 26 ASP N N 125.075 0.050 1 69 498 27 HIS H H 8.809 0.050 1 70 498 27 HIS CA C 56.913 0.106 1 71 498 27 HIS CB C 27.774 0.035 1 72 498 27 HIS N N 123.708 0.050 1 73 499 28 ASN H H 8.377 0.050 1 74 499 28 ASN CA C 53.766 0.050 1 75 499 28 ASN CB C 36.190 0.036 1 76 499 28 ASN N N 116.093 0.050 1 77 500 29 THR H H 6.555 0.050 1 78 500 29 THR CA C 58.717 0.050 1 79 500 29 THR CB C 65.106 4.368 1 80 500 29 THR N N 104.573 0.050 1 81 501 30 LYS H H 7.730 0.050 1 82 501 30 LYS CA C 54.686 0.142 1 83 501 30 LYS CB C 25.934 0.035 1 84 501 30 LYS N N 119.477 0.050 1 85 502 31 THR H H 7.257 0.050 1 86 502 31 THR CA C 57.691 0.106 1 87 502 31 THR CB C 69.998 0.106 1 88 502 31 THR N N 109.585 0.050 1 89 503 32 THR H H 8.017 0.050 1 90 503 32 THR CA C 57.939 0.050 1 91 503 32 THR CB C 69.008 0.177 1 92 503 32 THR N N 112.221 0.050 1 93 504 33 THR H H 9.208 0.050 1 94 504 33 THR CA C 57.196 0.035 1 95 504 33 THR CB C 66.992 0.071 1 96 504 33 THR N N 115.638 0.050 1 97 505 34 TRP H H 8.779 0.050 1 98 505 34 TRP CA C 55.499 0.318 1 99 505 34 TRP CB C 28.658 0.050 1 100 505 34 TRP N N 126.214 0.050 1 101 506 35 GLU H H 8.792 0.050 1 102 506 35 GLU CA C 54.190 0.071 1 103 506 35 GLU CB C 27.703 0.106 1 104 506 35 GLU N N 121.787 0.050 1 105 507 36 ASP H H 8.085 0.050 1 106 507 36 ASP CA C 48.037 0.050 1 107 507 36 ASP CB C 39.408 0.050 1 108 507 36 ASP N N 126.507 0.050 1 109 508 37 PRO CA C 60.909 0.050 1 110 509 38 ARG H H 8.312 0.050 1 111 509 38 ARG CA C 55.039 0.050 1 112 509 38 ARG CB C 27.385 0.050 1 113 509 38 ARG N N 119.152 0.050 1 stop_ save_