data_5186 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H, 13C, and 15N backbone assignment of the vascular endothelial growth factor receptor-binding domain ; _BMRB_accession_number 5186 _BMRB_flat_file_name bmr5186.str _Entry_type original _Submission_date 2001-10-19 _Accession_date 2001-10-22 _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 Pan Borlan . . 2 Fairbrother Wayne J. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 184 "13C chemical shifts" 260 "15N chemical shifts" 85 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2002-05-07 original author . stop_ loop_ _Related_BMRB_accession_number _Relationship 5185 'Assignments for vascular endothelial growth factor in complex with v107' 5198 'Assignments for v107 in complex with vascular endothelial growth factor' stop_ _Original_release_date 2002-05-07 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; 1H, 13C, and 15N Resonance Assignment of the Vascular Endothelial Growth Factor Receptor-binding Domain in Complex with a Receptor-blocking Peptide ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code 21880667 _PubMed_ID 11883783 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Pan Borlan . . 2 Fairbrother Wayne J. . stop_ _Journal_abbreviation 'J. Biomol. NMR' _Journal_volume 22 _Journal_issue 2 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 189 _Page_last 190 _Year 2002 _Details . loop_ _Keyword 'vascular endothelial growth factor' stop_ save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Fairbrother WJ, Champe MA, Christinger HW, Keyt BA, Starovasnik MA. 1H, 13C, and 15N backbone assignment and secondary structure of the receptor-binding domain of vascular endothelial growth factor. Protein Sci. 1997 Oct;6(10):2250-60. ; _Citation_title '1H, 13C, and 15N backbone assignment and secondary structure of the receptor-binding domain of vascular endothelial growth factor.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 9336848 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Fairbrother 'W J' J. . 2 Champe 'M A' A. . 3 Christinger 'H W' W. . 4 Keyt 'B A' A. . 5 Starovasnik 'M A' A. . stop_ _Journal_abbreviation 'Protein Sci.' _Journal_name_full 'Protein science : a publication of the Protein Society' _Journal_volume 6 _Journal_issue 10 _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 2250 _Page_last 2260 _Year 1997 _Details ; Nearly complete sequence-specific 1H, 13C, and 15N resonance assignments are reported for the backbone atoms of the receptor-binding domain of vascular endothelial growth factor (VEGF), a 23-kDa homodimeric protein that is a major regulator of both normal and pathological angiogenesis. The assignment strategy relied on the use of seven 3D triple-resonance experiments [HN(CO)CA, HNCA, HNCO, (HCA)CONH, HN(COCA)HA, HN(CA)HA, and CBCA-(CO)NH] and a 3D 15N-TOCSY-HSQC experiment recorded on a 0.5 mM (12 mg/mL) sample at 500 MHz, pH 7.0, 45 degrees C. Under these conditions, 15N relaxation data show that the protein has a rotational correlation time of 15.0 ns. Despite this unusually long correlation time, assignments were obtained for 94 of the 99 residues; 8 residues lack amide 1H and 15N assignments, presumably due to rapid exchange of the amide 1H with solvent under the experimental conditions used. The secondary structure of the protein was deduced from the chemical shift indices of the 1H alpha, 13C alpha, 13C beta, and 13CO nuclei, and from analysis of backbone NOEs observed in a 3D 15N-NOESY-HSQC spectrum. Two helices and a significant amount of beta-sheet structure were identified, in general agreement with the secondary structure found in a recently determined crystal structure of a similar VEGF construct [Muller YA et al., 1997, Proc Natl Acad Sci USA 94:7192-7197]. ; save_ ################################## # Molecular system description # ################################## save_system_VEGF _Saveframe_category molecular_system _Mol_system_name 'vascular endothelial growth factor receptor-binding domain dimer' _Abbreviation_common VEGF _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'VEGF subunit 1' $VEGF 'VEGF subunit 2' $VEGF stop_ _System_molecular_weight 23250.8 _System_physical_state native _System_oligomer_state dimer _System_paramagnetic no _System_thiol_state 'all disulfide bound' loop_ _Magnetic_equivalence_ID _Magnetically_equivalent_system_component 1 'VEGF subunit 1' 1 'VEGF subunit 2' stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_VEGF _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'vascular endothelial growth factor' _Name_variant VEGF(11-109) _Abbreviation_common VEGF _Molecular_mass 11625.4 _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 99 _Mol_residue_sequence ; HHEVVKFMDVYQRSYCHPIE TLVDIFQEYPDEIEYIFKPS CVPLMRCGGCCNDEGLECVP TEESNITMQIMRIKPHQGQH IGEMSFLQHNKCECRPKKD ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 11 HIS 2 12 HIS 3 13 GLU 4 14 VAL 5 15 VAL 6 16 LYS 7 17 PHE 8 18 MET 9 19 ASP 10 20 VAL 11 21 TYR 12 22 GLN 13 23 ARG 14 24 SER 15 25 TYR 16 26 CYS 17 27 HIS 18 28 PRO 19 29 ILE 20 30 GLU 21 31 THR 22 32 LEU 23 33 VAL 24 34 ASP 25 35 ILE 26 36 PHE 27 37 GLN 28 38 GLU 29 39 TYR 30 40 PRO 31 41 ASP 32 42 GLU 33 43 ILE 34 44 GLU 35 45 TYR 36 46 ILE 37 47 PHE 38 48 LYS 39 49 PRO 40 50 SER 41 51 CYS 42 52 VAL 43 53 PRO 44 54 LEU 45 55 MET 46 56 ARG 47 57 CYS 48 58 GLY 49 59 GLY 50 60 CYS 51 61 CYS 52 62 ASN 53 63 ASP 54 64 GLU 55 65 GLY 56 66 LEU 57 67 GLU 58 68 CYS 59 69 VAL 60 70 PRO 61 71 THR 62 72 GLU 63 73 GLU 64 74 SER 65 75 ASN 66 76 ILE 67 77 THR 68 78 MET 69 79 GLN 70 80 ILE 71 81 MET 72 82 ARG 73 83 ILE 74 84 LYS 75 85 PRO 76 86 HIS 77 87 GLN 78 88 GLY 79 89 GLN 80 90 HIS 81 91 ILE 82 92 GLY 83 93 GLU 84 94 MET 85 95 SER 86 96 PHE 87 97 LEU 88 98 GLN 89 99 HIS 90 100 ASN 91 101 LYS 92 102 CYS 93 103 GLU 94 104 CYS 95 105 ARG 96 106 PRO 97 107 LYS 98 108 LYS 99 109 ASP stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-01-28 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value BMRB 5198 VEGF 100.00 99 100.00 100.00 5.41e-66 PDB 1BJ1 "Vascular Endothelial Growth Factor In Complex With A Neutralizing Antibody" 100.00 102 100.00 100.00 5.80e-66 PDB 1CZ8 "Vascular Endothelial Growth Factor In Complex With An Affinity Matured Antibody" 94.95 94 100.00 100.00 5.27e-62 PDB 1FLT "Vegf In Complex With Domain 2 Of The Flt-1 Receptor" 98.99 98 100.00 100.00 4.65e-65 PDB 1KAT "Solution Structure Of A Phage-Derived Peptide Antagonist In Complex With Vascular Endothelial Growth Factor" 100.00 99 100.00 100.00 5.41e-66 PDB 1MJV "Disulfide Deficient Mutant Of Vascular Endothelial Growth Factor A (C51a And C60a)" 95.96 96 97.89 97.89 2.26e-61 PDB 1MKG "Disulfide Deficient Mutant Of Vascular Endothelial Growth Factor A (C57a And C102a)" 95.96 96 97.89 97.89 2.26e-61 PDB 1MKK "Disulfide Deficient Mutant Of Vascular Endothelial Growth Factor A (C61a And C104a)" 95.96 96 97.89 97.89 2.26e-61 PDB 1QTY "Vascular Endothelial Growth Factor In Complex With Domain 2 Of The Flt-1 Receptor" 100.00 102 100.00 100.00 5.80e-66 PDB 1TZH "Crystal Structure Of The Fab Yads1 Complexed With H-Vegf" 100.00 102 100.00 100.00 5.80e-66 PDB 1TZI "Crystal Structure Of The Fab Yads2 Complexed With H-Vegf" 100.00 102 100.00 100.00 5.80e-66 PDB 1VPF "Structure Of Human Vascular Endothelial Growth Factor" 100.00 102 100.00 100.00 5.80e-66 PDB 1VPP "Complex Between Vegf And A Receptor Blocking Peptide" 100.00 102 100.00 100.00 5.80e-66 PDB 2FJG "Structure Of The G6 Fab, A Phage Derived Fab Fragment, In Complex With Vegf" 100.00 102 100.00 100.00 5.80e-66 PDB 2FJH "Structure Of The B20-4 Fab, A Phage Derived Fab Fragment, In Complex With Vegf" 100.00 102 100.00 100.00 5.80e-66 PDB 2QR0 "Structure Of Vegf Complexed To A Fab Containing Tyr And Ser In The Cdrs" 97.98 97 100.00 100.00 3.66e-64 PDB 2VPF "Vascular Endothelial Growth Factor Refined To 1.93 Angstroms Resolution" 100.00 102 100.00 100.00 5.80e-66 PDB 3BDY "Dual Specific Bh1 Fab In Complex With Vegf" 100.00 102 100.00 100.00 5.80e-66 PDB 3P9W "Crystal Structure Of An Engineered Human Autonomous Vh Domain In Complex With Vegf" 100.00 106 100.00 100.00 3.06e-66 PDB 3QTK "The Crystal Structure Of Chemically Synthesized Vegf-A" 100.00 102 100.00 100.00 5.80e-66 PDB 3S1B "The Development Of Peptide-Based Tools For The Analysis Of Angiogenesis" 96.97 96 100.00 100.00 1.71e-63 PDB 3S1K "The Development Of Peptide-Based Tools For The Analysis Of Angiogenesis" 100.00 102 100.00 100.00 5.80e-66 PDB 3V2A "Vegfr-2VEGF-A Complex Structure" 100.00 134 100.00 100.00 5.35e-66 PDB 4GLN "Crystal Structure Of Chemically Synthesized Heterochiral {d-protein Antagonist Plus Vegf-a} Protein Complex In Space Group P21/" 100.00 102 100.00 100.00 5.80e-66 PDB 4GLS "Crystal Structure Of Chemically Synthesized Heterochiral {d-protein Antagonist Plus Vegf-a} Protein Complex In Space Group P21" 100.00 102 100.00 100.00 5.80e-66 PDB 4KZN "Crystal Structure Of Human Vegf-a Receptor Binding Domain" 100.00 104 100.00 100.00 7.51e-66 DBJ BAA78418 "vascular endothelial growth factor isoform VEGF165 [Homo sapiens]" 100.00 191 100.00 100.00 6.51e-66 DBJ BAB68520 "vascular endothelial growth factor [Felis catus]" 98.99 189 96.94 97.96 2.79e-62 DBJ BAD92722 "vascular endothelial growth factor variant [Homo sapiens]" 98.99 148 98.98 98.98 5.85e-65 DBJ BAG70136 "vascular endothelial growth factor isoform VEGF165 [Homo sapiens]" 100.00 191 100.00 100.00 6.51e-66 DBJ BAG70265 "vascular endothelial growth factor isoform VEGF165 [Homo sapiens]" 100.00 191 100.00 100.00 6.51e-66 EMBL CAA09179 "VEGF183 protein [Homo sapiens]" 100.00 209 100.00 100.00 3.19e-66 EMBL CAA44447 "vascular endothelial growth factor [Homo sapiens]" 100.00 191 100.00 100.00 6.51e-66 EMBL CAA57143 "unnamed protein product [Sus scrofa]" 98.99 190 97.96 97.96 5.28e-63 EMBL CAB82426 "vascular endothelial growth factor 188 [Canis lupus familiaris]" 98.99 214 96.94 97.96 2.18e-62 EMBL CAC19923 "vascular endothelial growth factor [Callithrix jacchus]" 95.96 124 98.95 98.95 2.11e-61 GB AAA35789 "vascular endothelial growth factor [Homo sapiens]" 100.00 191 100.00 100.00 6.51e-66 GB AAA36804 "vascular endothelial growth factor [Homo sapiens]" 100.00 215 100.00 100.00 2.28e-66 GB AAA36807 "vascular permeability factor precursor [Homo sapiens]" 100.00 215 100.00 100.00 2.28e-66 GB AAB47118 "simVEGF165 [Macaca fascicularis]" 100.00 191 100.00 100.00 6.51e-66 GB AAC63102 "vascular endothelial growth factor [Homo sapiens]" 100.00 254 100.00 100.00 9.73e-66 PRF 2105202A "vascular endothelial growth factor" 98.99 190 97.96 97.96 5.28e-63 REF NP_001003175 "vascular endothelial growth factor A isoform 1 precursor [Canis lupus familiaris]" 98.99 214 96.94 97.96 2.18e-62 REF NP_001009854 "vascular endothelial growth factor A precursor [Felis catus]" 98.99 189 96.94 97.96 2.79e-62 REF NP_001020537 "vascular endothelial growth factor A isoform a [Homo sapiens]" 100.00 412 100.00 100.00 9.44e-66 REF NP_001020538 "vascular endothelial growth factor A isoform c [Homo sapiens]" 100.00 389 100.00 100.00 5.62e-66 REF NP_001020539 "vascular endothelial growth factor A isoform d [Homo sapiens]" 100.00 371 100.00 100.00 8.37e-66 SP P15692 "RecName: Full=Vascular endothelial growth factor A; Short=VEGF-A; AltName: Full=Vascular permeability factor; Short=VPF; Flags:" 100.00 232 100.00 100.00 8.37e-66 SP P49151 "RecName: Full=Vascular endothelial growth factor A; Short=VEGF-A; AltName: Full=Vascular permeability factor; Short=VPF; Flags:" 98.99 190 97.96 97.96 5.28e-63 SP Q9MYV3 "RecName: Full=Vascular endothelial growth factor A; Short=VEGF-A; AltName: Full=Vascular permeability factor; Short=VPF; Flags:" 98.99 214 96.94 97.96 2.18e-62 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $VEGF 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 _Vector_name $VEGF 'recombinant technology' 'E. coli' . . . . 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 $VEGF 1.0 mM '[U-98% 13C; U-98% 15N]' stop_ save_ save_Sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $VEGF 2.0 mM '[U-98% 13C; U-98% 15N]' stop_ 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 800 _Details . save_ ####################### # Sample conditions # ####################### save_Cond_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 7.0 0.1 n/a temperature 318 0.2 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 . . . 1.0 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_ _Sample_label $Sample_1 $Sample_2 stop_ _Sample_conditions_label $Cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'VEGF subunit 1' _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 . 3 GLU HA H 4.30 0.02 1 2 . 3 GLU C C 175.90 0.20 1 3 . 3 GLU CA C 56.60 0.20 1 4 . 3 GLU CB C 31.30 0.20 1 5 . 4 VAL H H 8.05 0.02 1 6 . 4 VAL HA H 4.27 0.02 1 7 . 4 VAL C C 175.83 0.20 1 8 . 4 VAL CA C 61.66 0.20 1 9 . 4 VAL CB C 33.90 0.20 1 10 . 4 VAL N N 123.12 0.10 1 11 . 5 VAL H H 8.58 0.02 1 12 . 5 VAL HA H 4.07 0.02 1 13 . 5 VAL C C 175.89 0.20 1 14 . 5 VAL CA C 63.22 0.20 1 15 . 5 VAL CB C 32.30 0.20 1 16 . 5 VAL N N 128.73 0.10 1 17 . 6 LYS H H 8.90 0.02 1 18 . 6 LYS HA H 4.09 0.02 1 19 . 6 LYS C C 178.17 0.20 1 20 . 6 LYS CA C 56.85 0.20 1 21 . 6 LYS CB C 33.30 0.20 1 22 . 6 LYS N N 127.56 0.10 1 23 . 7 PHE H H 8.83 0.02 1 24 . 7 PHE HA H 4.62 0.02 1 25 . 7 PHE C C 176.31 0.20 1 26 . 7 PHE CA C 61.22 0.20 1 27 . 7 PHE CB C 40.10 0.20 1 28 . 7 PHE N N 122.81 0.10 1 29 . 8 MET H H 8.91 0.02 1 30 . 8 MET HA H 3.96 0.02 1 31 . 8 MET C C 178.39 0.20 1 32 . 8 MET CA C 57.87 0.20 1 33 . 8 MET CB C 31.60 0.20 1 34 . 8 MET N N 115.31 0.10 1 35 . 9 ASP H H 7.16 0.02 1 36 . 9 ASP HA H 4.49 0.02 1 37 . 9 ASP C C 178.04 0.20 1 38 . 9 ASP CA C 57.42 0.20 1 39 . 9 ASP CB C 41.30 0.20 1 40 . 9 ASP N N 118.56 0.10 1 41 . 10 VAL H H 7.95 0.02 1 42 . 10 VAL HA H 3.43 0.02 1 43 . 10 VAL C C 177.99 0.20 1 44 . 10 VAL CA C 66.73 0.20 1 45 . 10 VAL N N 119.59 0.10 1 46 . 11 TYR H H 8.65 0.02 1 47 . 11 TYR HA H 3.65 0.02 1 48 . 11 TYR C C 177.99 0.20 1 49 . 11 TYR CA C 62.19 0.20 1 50 . 11 TYR CB C 39.00 0.20 1 51 . 11 TYR N N 120.46 0.10 1 52 . 12 GLN H H 7.84 0.02 1 53 . 12 GLN HA H 3.94 0.02 1 54 . 12 GLN C C 178.57 0.20 1 55 . 12 GLN CA C 59.03 0.20 1 56 . 12 GLN CB C 29.30 0.20 1 57 . 12 GLN N N 115.56 0.10 1 58 . 13 ARG H H 7.87 0.02 1 59 . 13 ARG HA H 3.97 0.02 1 60 . 13 ARG C C 177.18 0.20 1 61 . 13 ARG CA C 58.54 0.20 1 62 . 13 ARG CB C 32.00 0.20 1 63 . 13 ARG N N 116.68 0.10 1 64 . 14 SER H H 7.78 0.02 1 65 . 14 SER HA H 4.28 0.02 1 66 . 14 SER C C 172.62 0.20 1 67 . 14 SER CA C 58.97 0.20 1 68 . 14 SER CB C 65.30 0.20 1 69 . 14 SER N N 112.37 0.10 1 70 . 15 TYR H H 6.79 0.02 1 71 . 15 TYR HA H 4.36 0.02 1 72 . 15 TYR C C 175.29 0.20 1 73 . 15 TYR CA C 59.66 0.20 1 74 . 15 TYR CB C 38.80 0.20 1 75 . 15 TYR N N 122.46 0.10 1 76 . 16 CYS H H 6.87 0.02 1 77 . 16 CYS HA H 4.65 0.02 1 78 . 16 CYS C C 171.64 0.20 1 79 . 16 CYS CA C 55.85 0.20 1 80 . 16 CYS CB C 39.00 0.20 1 81 . 16 CYS N N 126.65 0.10 1 82 . 17 HIS H H 9.20 0.02 1 83 . 17 HIS HA H 4.76 0.02 1 84 . 17 HIS C C 170.49 0.20 1 85 . 17 HIS CA C 55.54 0.20 1 86 . 17 HIS N N 124.53 0.10 1 87 . 18 PRO HA H 4.21 0.02 1 88 . 18 PRO C C 175.49 0.20 1 89 . 18 PRO CA C 63.30 0.20 1 90 . 19 ILE H H 9.00 0.02 1 91 . 19 ILE HA H 4.47 0.02 1 92 . 19 ILE C C 175.25 0.20 1 93 . 19 ILE CA C 59.45 0.20 1 94 . 19 ILE CB C 42.80 0.20 1 95 . 19 ILE N N 124.18 0.10 1 96 . 20 GLU H H 8.60 0.02 1 97 . 20 GLU HA H 4.15 0.02 1 98 . 20 GLU C C 175.33 0.20 1 99 . 20 GLU CA C 59.32 0.20 1 100 . 20 GLU N N 129.56 0.10 1 101 . 21 THR H H 9.36 0.02 1 102 . 21 THR HA H 4.25 0.02 1 103 . 21 THR C C 171.76 0.20 1 104 . 21 THR CA C 61.85 0.20 1 105 . 21 THR CB C 71.70 0.20 1 106 . 21 THR N N 127.06 0.10 1 107 . 22 LEU H H 8.58 0.02 1 108 . 22 LEU HA H 4.82 0.02 1 109 . 22 LEU C C 176.25 0.20 1 110 . 22 LEU CA C 53.39 0.20 1 111 . 22 LEU CB C 40.10 0.20 1 112 . 22 LEU N N 128.81 0.10 1 113 . 23 VAL H H 8.65 0.02 1 114 . 23 VAL HA H 4.07 0.02 1 115 . 23 VAL C C 175.51 0.20 1 116 . 23 VAL CA C 61.28 0.20 1 117 . 23 VAL CB C 35.10 0.20 1 118 . 23 VAL N N 125.43 0.10 1 119 . 24 ASP H H 8.55 0.02 1 120 . 24 ASP HA H 4.63 0.02 1 121 . 24 ASP C C 177.67 0.20 1 122 . 24 ASP CA C 55.02 0.20 1 123 . 24 ASP CB C 42.60 0.20 1 124 . 24 ASP N N 127.09 0.10 1 125 . 25 ILE H H 8.64 0.02 1 126 . 25 ILE HA H 3.41 0.02 1 127 . 25 ILE C C 178.14 0.20 1 128 . 25 ILE CA C 67.08 0.20 1 129 . 25 ILE CB C 37.90 0.20 1 130 . 25 ILE N N 127.68 0.10 1 131 . 26 PHE H H 8.53 0.02 1 132 . 26 PHE HA H 4.45 0.02 1 133 . 26 PHE C C 177.30 0.20 1 134 . 26 PHE CA C 59.86 0.20 1 135 . 26 PHE CB C 38.50 0.20 1 136 . 26 PHE N N 117.25 0.10 1 137 . 27 GLN H H 7.60 0.02 1 138 . 27 GLN HA H 4.10 0.02 1 139 . 27 GLN C C 177.43 0.20 1 140 . 27 GLN CA C 58.29 0.20 1 141 . 27 GLN CB C 29.20 0.20 1 142 . 27 GLN N N 116.25 0.10 1 143 . 28 GLU H H 7.24 0.02 1 144 . 28 GLU HA H 4.10 0.02 1 145 . 28 GLU C C 176.22 0.20 1 146 . 28 GLU CA C 57.07 0.20 1 147 . 28 GLU CB C 31.70 0.20 1 148 . 28 GLU N N 117.81 0.10 1 149 . 29 TYR H H 8.23 0.02 1 150 . 29 TYR HA H 4.87 0.02 1 151 . 29 TYR C C 173.48 0.20 1 152 . 29 TYR CA C 56.93 0.20 1 153 . 29 TYR N N 118.03 0.10 1 154 . 30 PRO HA H 4.59 0.02 1 155 . 30 PRO C C 177.94 0.20 1 156 . 30 PRO CA C 64.90 0.20 1 157 . 30 PRO CB C 32.50 0.20 1 158 . 31 ASP H H 8.51 0.02 1 159 . 31 ASP HA H 4.77 0.02 1 160 . 31 ASP C C 177.80 0.20 1 161 . 31 ASP CA C 54.92 0.20 1 162 . 31 ASP CB C 41.20 0.20 1 163 . 31 ASP N N 115.81 0.10 1 164 . 32 GLU H H 7.83 0.02 1 165 . 32 GLU HA H 4.84 0.02 1 166 . 32 GLU C C 176.70 0.20 1 167 . 32 GLU CA C 56.68 0.20 1 168 . 32 GLU CB C 29.10 0.20 1 169 . 32 GLU N N 121.99 0.10 1 170 . 33 ILE H H 7.37 0.02 1 171 . 33 ILE HA H 4.26 0.02 1 172 . 33 ILE C C 176.67 0.20 1 173 . 33 ILE CA C 62.60 0.20 1 174 . 33 ILE CB C 39.40 0.20 1 175 . 33 ILE N N 114.75 0.10 1 176 . 34 GLU H H 8.48 0.02 1 177 . 34 GLU HA H 4.11 0.02 1 178 . 34 GLU C C 175.79 0.20 1 179 . 34 GLU CA C 57.81 0.20 1 180 . 34 GLU CB C 30.50 0.20 1 181 . 34 GLU N N 120.68 0.10 1 182 . 35 TYR H H 7.66 0.02 1 183 . 35 TYR HA H 4.70 0.02 1 184 . 35 TYR C C 175.21 0.20 1 185 . 35 TYR CA C 57.74 0.20 1 186 . 35 TYR CB C 41.30 0.20 1 187 . 35 TYR N N 116.37 0.10 1 188 . 36 ILE H H 8.48 0.02 1 189 . 36 ILE HA H 4.19 0.02 1 190 . 36 ILE C C 174.58 0.20 1 191 . 36 ILE CA C 60.89 0.20 1 192 . 36 ILE CB C 40.40 0.20 1 193 . 36 ILE N N 118.12 0.10 1 194 . 37 PHE H H 8.16 0.02 1 195 . 37 PHE HA H 5.51 0.02 1 196 . 37 PHE C C 175.90 0.20 1 197 . 37 PHE CA C 56.96 0.20 1 198 . 37 PHE CB C 43.10 0.20 1 199 . 37 PHE N N 123.33 0.10 1 200 . 38 LYS H H 8.99 0.02 1 201 . 38 LYS HA H 4.60 0.02 1 202 . 38 LYS C C 174.84 0.20 1 203 . 38 LYS CA C 53.79 0.20 1 204 . 38 LYS N N 121.65 0.10 1 205 . 39 PRO HA H 4.77 0.02 1 206 . 39 PRO C C 173.90 0.20 1 207 . 39 PRO CA C 63.90 0.20 1 208 . 40 SER H H 8.48 0.02 1 209 . 40 SER HA H 4.27 0.02 1 210 . 40 SER C C 173.05 0.20 1 211 . 40 SER CA C 59.64 0.20 1 212 . 40 SER CB C 64.50 0.20 1 213 . 40 SER N N 109.06 0.10 1 214 . 41 CYS H H 8.13 0.02 1 215 . 41 CYS HA H 5.62 0.02 1 216 . 41 CYS C C 170.91 0.20 1 217 . 41 CYS CA C 56.03 0.20 1 218 . 41 CYS CB C 39.90 0.20 1 219 . 41 CYS N N 123.24 0.10 1 220 . 42 VAL H H 8.41 0.02 1 221 . 42 VAL HA H 4.74 0.02 1 222 . 42 VAL C C 172.82 0.20 1 223 . 42 VAL CA C 57.05 0.20 1 224 . 42 VAL N N 109.69 0.10 1 225 . 43 PRO HA H 4.91 0.02 1 226 . 43 PRO C C 175.34 0.20 1 227 . 43 PRO CA C 61.00 0.20 1 228 . 43 PRO CB C 30.80 0.20 1 229 . 44 LEU H H 9.01 0.02 1 230 . 44 LEU HA H 4.68 0.02 1 231 . 44 LEU C C 176.46 0.20 1 232 . 44 LEU CA C 52.83 0.20 1 233 . 44 LEU CB C 47.40 0.20 1 234 . 44 LEU N N 124.31 0.10 1 235 . 45 MET H H 8.62 0.02 1 236 . 45 MET HA H 4.77 0.02 1 237 . 45 MET C C 176.77 0.20 1 238 . 45 MET CA C 54.05 0.20 1 239 . 45 MET CB C 31.60 0.20 1 240 . 45 MET N N 121.25 0.10 1 241 . 46 ARG H H 8.65 0.02 1 242 . 46 ARG HA H 4.54 0.02 1 243 . 46 ARG C C 176.29 0.20 1 244 . 46 ARG CA C 52.32 0.20 1 245 . 46 ARG CB C 34.70 0.20 1 246 . 46 ARG N N 125.54 0.10 1 247 . 47 CYS H H 12.11 0.02 1 248 . 47 CYS HA H 4.29 0.02 1 249 . 47 CYS C C 175.18 0.20 1 250 . 47 CYS CA C 57.15 0.20 1 251 . 47 CYS CB C 39.80 0.20 1 252 . 47 CYS N N 125.81 0.10 1 253 . 48 GLY H H 8.40 0.02 1 254 . 48 GLY HA2 H 4.28 0.02 2 255 . 48 GLY HA3 H 3.61 0.02 2 256 . 48 GLY C C 170.56 0.20 1 257 . 48 GLY CA C 44.55 0.20 1 258 . 48 GLY N N 111.97 0.10 1 259 . 49 GLY H H 8.27 0.02 1 260 . 49 GLY HA2 H 4.53 0.02 2 261 . 49 GLY HA3 H 3.41 0.02 2 262 . 49 GLY C C 173.95 0.20 1 263 . 49 GLY CA C 44.98 0.20 1 264 . 49 GLY N N 103.94 0.10 1 265 . 50 CYS H H 8.09 0.02 1 266 . 50 CYS HA H 5.22 0.02 1 267 . 50 CYS C C 175.09 0.20 1 268 . 50 CYS CA C 53.17 0.20 1 269 . 50 CYS N N 111.56 0.10 1 270 . 52 ASN HA H 4.59 0.02 1 271 . 52 ASN C C 173.60 0.20 1 272 . 52 ASN CA C 55.60 0.20 1 273 . 52 ASN CB C 38.50 0.20 1 274 . 53 ASP H H 7.17 0.02 1 275 . 53 ASP HA H 4.71 0.02 1 276 . 53 ASP C C 176.40 0.20 1 277 . 53 ASP CA C 54.40 0.20 1 278 . 53 ASP N N 115.96 0.10 1 279 . 55 GLY H H 9.08 0.02 1 280 . 55 GLY HA2 H 4.14 0.02 2 281 . 55 GLY HA3 H 3.80 0.02 2 282 . 55 GLY C C 174.00 0.20 1 283 . 55 GLY N N 108.28 0.10 1 284 . 56 LEU H H 7.49 0.02 1 285 . 56 LEU HA H 4.96 0.02 1 286 . 56 LEU C C 175.16 0.20 1 287 . 56 LEU CA C 53.50 0.20 1 288 . 56 LEU CB C 45.80 0.20 1 289 . 56 LEU N N 119.62 0.10 1 290 . 57 GLU H H 9.16 0.02 1 291 . 57 GLU HA H 4.62 0.02 1 292 . 57 GLU C C 173.84 0.20 1 293 . 57 GLU CA C 53.91 0.20 1 294 . 57 GLU CB C 33.40 0.20 1 295 . 57 GLU N N 118.71 0.10 1 296 . 58 CYS H H 8.65 0.02 1 297 . 58 CYS HA H 4.76 0.02 1 298 . 58 CYS C C 173.89 0.20 1 299 . 58 CYS CA C 54.72 0.20 1 300 . 58 CYS CB C 36.30 0.20 1 301 . 58 CYS N N 123.06 0.10 1 302 . 59 VAL H H 8.93 0.02 1 303 . 59 VAL HA H 4.82 0.02 1 304 . 59 VAL C C 172.72 0.20 1 305 . 59 VAL CA C 57.97 0.20 1 306 . 59 VAL N N 124.18 0.10 1 307 . 60 PRO HA H 5.01 0.02 1 308 . 60 PRO C C 178.48 0.20 1 309 . 60 PRO CA C 62.50 0.20 1 310 . 61 THR H H 8.45 0.02 1 311 . 61 THR HA H 4.36 0.02 1 312 . 61 THR C C 175.17 0.20 1 313 . 61 THR CA C 61.24 0.20 1 314 . 61 THR CB C 69.10 0.20 1 315 . 61 THR N N 111.06 0.10 1 316 . 62 GLU H H 7.20 0.02 1 317 . 62 GLU HA H 4.54 0.02 1 318 . 62 GLU C C 175.06 0.20 1 319 . 62 GLU CA C 57.16 0.20 1 320 . 62 GLU CB C 34.40 0.20 1 321 . 62 GLU N N 120.90 0.10 1 322 . 63 GLU H H 8.82 0.02 1 323 . 63 GLU HA H 5.43 0.02 1 324 . 63 GLU C C 175.01 0.20 1 325 . 63 GLU CA C 55.24 0.20 1 326 . 63 GLU CB C 34.30 0.20 1 327 . 63 GLU N N 125.81 0.10 1 328 . 64 SER H H 8.82 0.02 1 329 . 64 SER HA H 4.69 0.02 1 330 . 64 SER C C 172.55 0.20 1 331 . 64 SER CA C 57.96 0.20 1 332 . 64 SER CB C 65.40 0.20 1 333 . 64 SER N N 113.59 0.10 1 334 . 65 ASN H H 8.52 0.02 1 335 . 65 ASN HA H 5.75 0.02 1 336 . 65 ASN C C 174.97 0.20 1 337 . 65 ASN CA C 53.07 0.20 1 338 . 65 ASN CB C 41.90 0.20 1 339 . 65 ASN N N 117.96 0.10 1 340 . 66 ILE H H 9.35 0.02 1 341 . 66 ILE HA H 4.68 0.02 1 342 . 66 ILE C C 172.86 0.20 1 343 . 66 ILE CA C 59.74 0.20 1 344 . 66 ILE CB C 42.60 0.20 1 345 . 66 ILE N N 122.65 0.10 1 346 . 67 THR H H 8.49 0.02 1 347 . 67 THR HA H 5.18 0.02 1 348 . 67 THR C C 173.48 0.20 1 349 . 67 THR CA C 62.18 0.20 1 350 . 67 THR CB C 69.80 0.20 1 351 . 67 THR N N 123.15 0.10 1 352 . 68 MET H H 9.21 0.02 1 353 . 68 MET HA H 5.09 0.02 1 354 . 68 MET C C 174.63 0.20 1 355 . 68 MET CA C 54.61 0.20 1 356 . 68 MET CB C 37.70 0.20 1 357 . 68 MET N N 123.68 0.10 1 358 . 69 GLN H H 8.15 0.02 1 359 . 69 GLN HA H 4.79 0.02 1 360 . 69 GLN C C 173.16 0.20 1 361 . 69 GLN CA C 55.86 0.20 1 362 . 69 GLN CB C 30.10 0.20 1 363 . 69 GLN N N 121.21 0.10 1 364 . 70 ILE H H 8.26 0.02 1 365 . 70 ILE HA H 4.67 0.02 1 366 . 70 ILE C C 176.46 0.20 1 367 . 70 ILE CA C 57.73 0.20 1 368 . 70 ILE CB C 41.80 0.20 1 369 . 70 ILE N N 119.97 0.10 1 370 . 71 MET H H 8.91 0.02 1 371 . 71 MET HA H 4.59 0.02 1 372 . 71 MET C C 174.08 0.20 1 373 . 71 MET CA C 55.98 0.20 1 374 . 71 MET CB C 34.10 0.20 1 375 . 71 MET N N 126.62 0.10 1 376 . 72 ARG H H 8.80 0.02 1 377 . 72 ARG HA H 4.67 0.02 1 378 . 72 ARG C C 175.44 0.20 1 379 . 72 ARG CA C 55.43 0.20 1 380 . 72 ARG CB C 32.50 0.20 1 381 . 72 ARG N N 127.62 0.10 1 382 . 73 ILE H H 9.20 0.02 1 383 . 73 ILE HA H 4.37 0.02 1 384 . 73 ILE C C 175.21 0.20 1 385 . 73 ILE CA C 61.39 0.20 1 386 . 73 ILE CB C 40.70 0.20 1 387 . 73 ILE N N 126.23 0.10 1 388 . 74 LYS H H 8.40 0.02 1 389 . 74 LYS HA H 4.96 0.02 1 390 . 74 LYS C C 174.55 0.20 1 391 . 74 LYS CA C 52.95 0.20 1 392 . 74 LYS N N 128.56 0.10 1 393 . 76 HIS HA H 4.25 0.02 1 394 . 76 HIS C C 175.20 0.20 1 395 . 76 HIS CA C 58.30 0.20 1 396 . 76 HIS CB C 29.30 0.20 1 397 . 77 GLN H H 8.26 0.02 1 398 . 77 GLN HA H 4.61 0.02 1 399 . 77 GLN C C 175.80 0.20 1 400 . 77 GLN CA C 56.60 0.20 1 401 . 77 GLN CB C 31.50 0.20 1 402 . 77 GLN N N 118.77 0.10 1 403 . 78 GLY H H 8.16 0.02 1 404 . 78 GLY HA2 H 4.22 0.02 2 405 . 78 GLY HA3 H 3.94 0.02 2 406 . 78 GLY C C 172.06 0.20 1 407 . 78 GLY CA C 45.71 0.20 1 408 . 78 GLY N N 107.78 0.10 1 409 . 79 GLN H H 8.14 0.02 1 410 . 79 GLN HA H 5.24 0.02 1 411 . 79 GLN C C 174.60 0.20 1 412 . 79 GLN CA C 54.30 0.20 1 413 . 79 GLN CB C 32.60 0.20 1 414 . 79 GLN N N 116.81 0.10 1 415 . 80 HIS H H 8.38 0.02 1 416 . 80 HIS HA H 4.74 0.02 1 417 . 80 HIS C C 173.60 0.20 1 418 . 80 HIS CA C 56.00 0.20 1 419 . 80 HIS N N 118.96 0.10 1 420 . 81 ILE H H 8.49 0.02 1 421 . 81 ILE HA H 4.30 0.02 1 422 . 81 ILE C C 175.70 0.20 1 423 . 81 ILE CA C 61.20 0.20 1 424 . 81 ILE CB C 37.80 0.20 1 425 . 81 ILE N N 124.33 0.10 1 426 . 82 GLY H H 8.61 0.02 1 427 . 82 GLY HA2 H 4.56 0.02 2 428 . 82 GLY HA3 H 3.60 0.02 2 429 . 82 GLY C C 171.76 0.20 1 430 . 82 GLY CA C 44.16 0.20 1 431 . 82 GLY N N 115.93 0.10 1 432 . 83 GLU H H 8.23 0.02 1 433 . 83 GLU HA H 4.75 0.02 1 434 . 83 GLU C C 176.75 0.20 1 435 . 83 GLU CA C 56.30 0.20 1 436 . 83 GLU CB C 31.50 0.20 1 437 . 83 GLU N N 119.14 0.10 1 438 . 84 MET H H 9.03 0.02 1 439 . 84 MET HA H 4.46 0.02 1 440 . 84 MET C C 173.73 0.20 1 441 . 84 MET CA C 55.47 0.20 1 442 . 84 MET CB C 38.10 0.20 1 443 . 84 MET N N 123.24 0.10 1 444 . 85 SER H H 7.90 0.02 1 445 . 85 SER HA H 5.40 0.02 1 446 . 85 SER C C 173.83 0.20 1 447 . 85 SER CA C 57.20 0.20 1 448 . 85 SER CB C 62.20 0.20 1 449 . 85 SER N N 116.50 0.10 1 450 . 86 PHE H H 9.27 0.02 1 451 . 86 PHE HA H 4.91 0.02 1 452 . 86 PHE C C 174.34 0.20 1 453 . 86 PHE CA C 57.00 0.20 1 454 . 86 PHE CB C 45.90 0.20 1 455 . 86 PHE N N 122.71 0.10 1 456 . 87 LEU H H 8.96 0.02 1 457 . 87 LEU HA H 4.82 0.02 1 458 . 87 LEU C C 176.96 0.20 1 459 . 87 LEU CA C 55.39 0.20 1 460 . 87 LEU CB C 44.40 0.20 1 461 . 87 LEU N N 123.15 0.10 1 462 . 88 GLN H H 9.41 0.02 1 463 . 88 GLN HA H 4.70 0.02 1 464 . 88 GLN C C 174.06 0.20 1 465 . 88 GLN CA C 54.66 0.20 1 466 . 88 GLN CB C 33.10 0.20 1 467 . 88 GLN N N 122.71 0.10 1 468 . 89 HIS H H 8.48 0.02 1 469 . 89 HIS HA H 5.16 0.02 1 470 . 89 HIS C C 174.34 0.20 1 471 . 89 HIS CA C 52.86 0.20 1 472 . 89 HIS CB C 34.00 0.20 1 473 . 89 HIS N N 120.80 0.10 1 474 . 90 ASN H H 8.60 0.02 1 475 . 90 ASN HA H 4.69 0.02 1 476 . 90 ASN C C 175.01 0.20 1 477 . 90 ASN CA C 54.23 0.20 1 478 . 90 ASN CB C 40.70 0.20 1 479 . 90 ASN N N 119.37 0.10 1 480 . 91 LYS H H 7.59 0.02 1 481 . 91 LYS HA H 4.39 0.02 1 482 . 91 LYS C C 175.29 0.20 1 483 . 91 LYS CA C 56.50 0.20 1 484 . 91 LYS CB C 37.60 0.20 1 485 . 91 LYS N N 117.65 0.10 1 486 . 92 CYS H H 8.80 0.02 1 487 . 92 CYS HA H 5.46 0.02 1 488 . 92 CYS C C 173.21 0.20 1 489 . 92 CYS CA C 54.47 0.20 1 490 . 92 CYS CB C 48.50 0.20 1 491 . 92 CYS N N 121.90 0.10 1 492 . 93 GLU H H 9.31 0.02 1 493 . 93 GLU HA H 4.57 0.02 1 494 . 93 GLU C C 174.24 0.20 1 495 . 93 GLU CA C 55.17 0.20 1 496 . 93 GLU CB C 35.50 0.20 1 497 . 93 GLU N N 121.12 0.10 1 498 . 94 CYS H H 8.44 0.02 1 499 . 94 CYS HA H 5.23 0.02 1 500 . 94 CYS C C 174.65 0.20 1 501 . 94 CYS CA C 56.96 0.20 1 502 . 94 CYS CB C 44.80 0.20 1 503 . 94 CYS N N 120.12 0.10 1 504 . 95 ARG H H 9.23 0.02 1 505 . 95 ARG HA H 4.92 0.02 1 506 . 95 ARG C C 173.43 0.20 1 507 . 95 ARG CA C 53.20 0.20 1 508 . 95 ARG N N 126.12 0.10 1 509 . 96 PRO HA H 4.66 0.02 1 510 . 96 PRO C C 177.24 0.20 1 511 . 96 PRO CA C 63.60 0.20 1 512 . 96 PRO CB C 32.40 0.20 1 513 . 97 LYS H H 8.11 0.02 1 514 . 97 LYS HA H 4.07 0.02 1 515 . 97 LYS C C 176.43 0.20 1 516 . 97 LYS CA C 57.25 0.20 1 517 . 97 LYS CB C 34.00 0.20 1 518 . 97 LYS N N 124.06 0.10 1 519 . 98 LYS H H 8.38 0.02 1 520 . 98 LYS HA H 4.31 0.02 1 521 . 98 LYS C C 175.33 0.20 1 522 . 98 LYS CA C 56.59 0.20 1 523 . 98 LYS CB C 33.70 0.20 1 524 . 98 LYS N N 123.59 0.10 1 525 . 99 ASP H H 7.84 0.02 1 526 . 99 ASP HA H 4.36 0.02 1 527 . 99 ASP C C 180.79 0.20 1 528 . 99 ASP CA C 56.20 0.20 1 529 . 99 ASP N N 127.24 0.10 1 stop_ save_