data_5896 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Unusual Ca2+-calmodulin binding interactions of the microtubule-associated protein F-STOP ; _BMRB_accession_number 5896 _BMRB_flat_file_name bmr5896.str _Entry_type original _Submission_date 2003-08-05 _Accession_date 2003-08-05 _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 Bouvier Denis . . 2 Vanhaverbeke Cecile . . 3 Simorre Jean-Pierre . . 4 Gans Pierre . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 152 "13C chemical shifts" 153 "15N chemical shifts" 152 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2004-02-11 original author . stop_ _Original_release_date 2004-02-11 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Unusual Ca(2+)-calmodulin Binding Interactions of the Microtubule-associated Protein F-STOP ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code 22878800 _PubMed_ID 14516200 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Bouvier Denis . . 2 Vanhaverbeke Cecile . . 3 Simorre Jean-Pierre . . 4 Arlaud Gerard . . 5 Bally Isabelle . . 6 Forge Vincent . . 7 Margolis Robert L. . 8 Gans Pierre . . 9 Kleman Jean-Philippe . . stop_ _Journal_abbreviation Biochemistry _Journal_volume 42 _Journal_issue 39 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 11484 _Page_last 11493 _Year 2003 _Details . save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Roberts DM, Crea R, Malecha M, Alvarado-Urbina G, Chiarello RH, Watterson DM. Chemical synthesis and expression of a calmodulin gene designed for site-specific mutagenesis. Biochemistry. 1985 24(19):5090-8. ; _Citation_title 'Chemical synthesis and expression of a calmodulin gene designed for site-specific mutagenesis.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 3000422 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Roberts 'D. M.' M. . 2 Crea R. . . 3 Malecha M. . . 4 Alvarado-Urbina G. . . 5 Chiarello 'R. H.' H. . 6 Watterson 'D. M.' M. . stop_ _Journal_abbreviation Biochemistry _Journal_name_full Biochemistry _Journal_volume 24 _Journal_issue 19 _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 5090 _Page_last 5098 _Year 1985 _Details ; A gene coding for a calmodulin was synthesized and expressed in Escherichia coli. The gene was produced by the enzymatic ligation of 61 chemically synthesized DNA fragments. The gene possesses 27 unique, regularly spaced, restriction endonuclease cleavage sites to facilitate gene mutagenesis by the replacement of specific gene segments with synthetic double-stranded DNA. An expression vector containing the calmodulin gene was used to transform E. coli. Purification and characterization of calmodulin (VU-1 calmodulin) expressed by these transformants showed that it lacks two posttranslational modifications: an amino-terminal blocking group and N epsilon, N epsilon, N epsilon-trimethyllysine at position 115. The cyclic nucleotide phosphodiesterase activator properties of VU-1, higher plant, and vertebrate calmodulins were not statistically different. However, VU-1 calmodulin was found to activate nicotinamide adenine dinucleotide (NAD) kinase to a maximal level that was at least 3-fold higher than that found with higher plant and vertebrate calmodulins. This higher level of activation is also characteristic of calmodulins from Dictyostelium discoideum and Chlamydomonas reinhardtii [Roberts, D. M., Burgess, W. H., & Watterson, D. M. (1984) Plant Physiol. 75, 796-798; Marshak, D. R., Clarke, M., Roberts, D. M., & Watterson, D. M. (1984) Biochemistry 23, 2891-2899]. The only common feature among Dictyostelium, Chlamydomonas, and VU-1 calmodulins not found in higher plant and vertebrate calmodulins is an unmethylated lysine at position 115. The results indicate that the lack of methylation of lysine-115 may contribute to the maximal level of NAD kinase activation.(ABSTRACT TRUNCATED AT 250 WORDS) ; save_ save_ref_2 _Saveframe_category citation _Citation_full ; Bosc C, Frank R, Denarier E, Ronjat M, Schweitzer A, Wehland J, Job D. Identification of novel bifunctional calmodulin-binding and microtubule-stabilizing motifs in STOP proteins. J Biol Chem. 2001 276(33):30904-13. ; _Citation_title 'Identification of novel bifunctional calmodulin-binding and microtubule-stabilizing motifs in STOP proteins.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11413126 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Bosc C. . . 2 Frank R. . . 3 Denarier E. . . 4 Ronjat M. . . 5 Schweitzer A. . . 6 Wehland J. . . 7 Job D. . . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'The Journal of biological chemistry' _Journal_volume 276 _Journal_issue 33 _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 30904 _Page_last 30913 _Year 2001 _Details ; Although microtubules are intrinsically labile tubulin assemblies, many cell types contain stable polymers, resisting depolymerizing conditions such as exposure to the cold or the drug nocodazole. This microtubule stabilization is largely due to polymer association with STOP proteins. There are several STOP variants, some with capacity to induce microtubule resistance to both the cold and nocodazole, others with microtubule cold stabilizing activity only. These microtubule-stabilizing effects of STOP proteins are inhibited by calmodulin and we now demonstrate that they are determined by two distinct kinds of repeated modular sequences (Mn and Mc), both containing a calmodulin-binding peptide, but displaying different microtubule stabilizing activities. Mn modules induce microtubule resistance to both the cold and nocodazole when expressed in cells. Mc modules, which correspond to the STOP central repeats, have microtubule cold stabilizing activity only. Mouse neuronal STOPs, which induce both cold and drug resistance in cellular microtubules, contain three Mn modules and four Mc modules. Compared with neuronal STOPs, the non-neuronal F-STOP lacks multiple Mn modules and this corresponds with an inability to induce nocodazole resistance. STOP modules represent novel bifunctional calmodulin-binding and microtubule-stabilizing sequences that may be essential for the generation of the different patterns of microtubule stabilization observed in cells. ; save_ ################################## # Molecular system description # ################################## save_CAM-STP23_complex _Saveframe_category molecular_system _Mol_system_name 'VU-1 calmodulin/F-stop complex' _Abbreviation_common 'CAM/STP23 complex' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label calmodulin $CAM 'F-STOP Mc module' $STP23 'CALCIUM (II) ION' $CA stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state heterodimer _System_paramagnetic no _System_thiol_state 'not present' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_CAM _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common calmodulin _Name_variant VU-CAM _Abbreviation_common CAM _Molecular_mass 18522 _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 163 _Mol_residue_sequence ; MSHHHHHHSMDIEFMADQLT DEQIAEFKEAFSLFDKDGDG TITTKELGTVMRSLGQNPTE AELQDMINEVDADGNGTIDF PEFLNLMARKMKDTDSEEEL KEAFRVFDKDGNGFISAAEL RHVMTNLGEKLTDEEVDEMI READVDGDGQVNYEEFVQVM MAK ; loop_ _Residue_seq_code _Residue_label 1 MET 2 SER 3 HIS 4 HIS 5 HIS 6 HIS 7 HIS 8 HIS 9 SER 10 MET 11 ASP 12 ILE 13 GLU 14 PHE 15 MET 16 ALA 17 ASP 18 GLN 19 LEU 20 THR 21 ASP 22 GLU 23 GLN 24 ILE 25 ALA 26 GLU 27 PHE 28 LYS 29 GLU 30 ALA 31 PHE 32 SER 33 LEU 34 PHE 35 ASP 36 LYS 37 ASP 38 GLY 39 ASP 40 GLY 41 THR 42 ILE 43 THR 44 THR 45 LYS 46 GLU 47 LEU 48 GLY 49 THR 50 VAL 51 MET 52 ARG 53 SER 54 LEU 55 GLY 56 GLN 57 ASN 58 PRO 59 THR 60 GLU 61 ALA 62 GLU 63 LEU 64 GLN 65 ASP 66 MET 67 ILE 68 ASN 69 GLU 70 VAL 71 ASP 72 ALA 73 ASP 74 GLY 75 ASN 76 GLY 77 THR 78 ILE 79 ASP 80 PHE 81 PRO 82 GLU 83 PHE 84 LEU 85 ASN 86 LEU 87 MET 88 ALA 89 ARG 90 LYS 91 MET 92 LYS 93 ASP 94 THR 95 ASP 96 SER 97 GLU 98 GLU 99 GLU 100 LEU 101 LYS 102 GLU 103 ALA 104 PHE 105 ARG 106 VAL 107 PHE 108 ASP 109 LYS 110 ASP 111 GLY 112 ASN 113 GLY 114 PHE 115 ILE 116 SER 117 ALA 118 ALA 119 GLU 120 LEU 121 ARG 122 HIS 123 VAL 124 MET 125 THR 126 ASN 127 LEU 128 GLY 129 GLU 130 LYS 131 LEU 132 THR 133 ASP 134 GLU 135 GLU 136 VAL 137 ASP 138 GLU 139 MET 140 ILE 141 ARG 142 GLU 143 ALA 144 ASP 145 VAL 146 ASP 147 GLY 148 ASP 149 GLY 150 GLN 151 VAL 152 ASN 153 TYR 154 GLU 155 GLU 156 PHE 157 VAL 158 GLN 159 VAL 160 MET 161 MET 162 ALA 163 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2014-10-26 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 GB ERE78236 "microtubule-associated protein 6 [Cricetulus griseus]" 65.22 130 100.00 100.00 7.60e-01 stop_ save_ save_STP23 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'F-STOP Mc module' _Abbreviation_common STP23 _Molecular_mass 2739 _Mol_thiol_state 'not present' _Details . _Residue_count 23 _Mol_residue_sequence ; QRDTRRKAGPAWMVTRTEGH EEK ; loop_ _Residue_seq_code _Residue_label 1 GLN 2 ARG 3 ASP 4 THR 5 ARG 6 ARG 7 LYS 8 ALA 9 GLY 10 PRO 11 ALA 12 TRP 13 MET 14 VAL 15 THR 16 ARG 17 THR 18 GLU 19 GLY 20 HIS 21 GLU 22 GLU 23 LYS stop_ _Sequence_homology_query_date 2008-08-19 _Sequence_homology_query_revised_last_date 2007-10-08 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 REF NP_058900.1 'microtubule-associated protein 6[Rattus norvegicus]' 2.42 952 100 100 2e-05 REF NP_058900.1 'microtubule-associated protein 6[Rattus norvegicus]' 2.42 952 100 100 2e-05 REF NP_058900.1 'microtubule-associated protein 6[Rattus norvegicus]' 2.42 952 100 100 5e-06 REF NP_058900.1 'microtubule-associated protein 6[Rattus norvegicus]' 2.42 952 100 100 5e-06 PRF 2208500A 'microtubule-stabilizing protein' 2.42 952 100 100 5e-06 REF NP_001036820.2 'microtubule-associated protein 6isoform 3 [Mus musculus]' 7.52 306 100 100 2e-05 GenBank EDM18414.1 'rCG39515, isoform CRA_a [Rattusnorvegicus]' 2.42 952 100 100 5e-06 GenBank EDM18414.1 'rCG39515, isoform CRA_a [Rattusnorvegicus]' 2.42 952 100 100 2e-05 GenBank EDM18414.1 'rCG39515, isoform CRA_a [Rattusnorvegicus]' 2.42 952 100 100 5e-06 GenBank EDM18414.1 'rCG39515, isoform CRA_a [Rattusnorvegicus]' 2.42 952 100 100 5e-06 EMBL CAA05555.1 'E-STOP protein [Rattus norvegicus]' 3.48 660 100 100 2e-05 GenBank EDM18415.1 'rCG39515, isoform CRA_b [Rattusnorvegicus]' 3.48 660 100 100 5e-06 EMBL CAA05555.1 'E-STOP protein [Rattus norvegicus]' 3.48 660 100 100 5e-06 EMBL CAA05555.1 'E-STOP protein [Rattus norvegicus]' 3.48 660 100 100 2e-05 EMBL CAA05555.1 'E-STOP protein [Rattus norvegicus]' 3.48 660 100 100 5e-06 EMBL CAA05555.1 'E-STOP protein [Rattus norvegicus]' 3.48 660 100 100 5e-06 stop_ save_ ############# # Ligands # ############# save_CA _Saveframe_category ligand _Mol_type "non-polymer (NON-POLYMER)" _Name_common 'CALCIUM ION' _BMRB_code CA _PDB_code CA _Molecular_mass 40.078 _Mol_charge 2 _Mol_paramagnetic . _Mol_aromatic no _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons CA CA CA . 2 . ? 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 $CAM rabbit 9986 Eukaryota Metazoa Oryctolagus cuniculus $STP23 'house mouse' 10090 Eukaryota Metazoa mus musculus 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 _Details $STP23 'chemical synthesis' . . . . . . $CAM 'recombinant technology' 'E. coli' . . BL21(DE3) . 'Synthetic construct for VU-1 calmodulin described in ref 1' 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 $CAM 0.7 mM '[U-97% 13C; U-98% 15N]' $STP23 0.7 mM . stop_ save_ ############################ # Computer software used # ############################ save_FELIX _Saveframe_category software _Name FELIX _Version 2000 loop_ _Task 'raw spectral data processing' 'peak picking' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model Inova _Field_strength 600 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model Inova _Field_strength 800 _Details . save_ ############################# # NMR applied experiments # ############################# save_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC' _Sample_label $sample_1 save_ save_HNCA_2 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA _Sample_label $sample_1 save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-15N HSQC' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_Condition_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 7.7 0.05 n/a temperature 300 0.5 K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference _Saveframe_category chemical_shift_reference _Details . loop_ _Mol_common_name _Atom_type _Atom_isotope_number _Atom_group _Chem_shift_units _Chem_shift_value _Reference_method _Reference_type _External_reference_sample_geometry _External_reference_location _External_reference_axis _Indirect_shift_ratio DSS H 1 'methyl protons' ppm 0.0 internal direct . . . 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 stop_ _Sample_conditions_label $Condition_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name calmodulin _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 . 10 MET H H 8.36 0.02 1 2 . 10 MET CA C 56.3 0.2 1 3 . 10 MET N N 121.7 0.2 1 4 . 11 ASP H H 8.17 0.02 1 5 . 11 ASP CA C 54.9 0.2 1 6 . 11 ASP N N 121.3 0.2 1 7 . 12 ILE H H 7.83 0.02 1 8 . 12 ILE CA C 61.7 0.2 1 9 . 12 ILE N N 120.1 0.2 1 10 . 13 GLU H H 8.28 0.02 1 11 . 13 GLU CA C 57.4 0.2 1 12 . 13 GLU N N 123.9 0.2 1 13 . 14 PHE H H 8.05 0.02 1 14 . 14 PHE CA C 58.3 0.2 1 15 . 14 PHE N N 120.9 0.2 1 16 . 15 MET H H 7.96 0.02 1 17 . 15 MET CA C 55.9 0.2 1 18 . 15 MET N N 121.8 0.2 1 19 . 16 ALA H H 8.08 0.02 1 20 . 16 ALA CA C 54.1 0.2 1 21 . 16 ALA N N 123.8 0.2 1 22 . 17 ASP H H 8.01 0.02 1 23 . 17 ASP CA C 55.3 0.2 1 24 . 17 ASP N N 116.5 0.2 1 25 . 18 GLN H H 7.84 0.02 1 26 . 18 GLN CA C 55.9 0.2 1 27 . 18 GLN N N 117.9 0.2 1 28 . 19 LEU H H 7.66 0.02 1 29 . 19 LEU CA C 54.6 0.2 1 30 . 19 LEU N N 121.2 0.2 1 31 . 20 THR H H 8.68 0.02 1 32 . 20 THR CA C 60.9 0.2 1 33 . 20 THR N N 112.9 0.2 1 34 . 21 ASP H H 8.87 0.02 1 35 . 21 ASP CA C 58.2 0.2 1 36 . 21 ASP N N 120.9 0.2 1 37 . 22 GLU H H 8.63 0.02 1 38 . 22 GLU CA C 60.4 0.2 1 39 . 22 GLU N N 120.4 0.2 1 40 . 23 GLN H H 7.68 0.02 1 41 . 23 GLN CA C 59.0 0.2 1 42 . 23 GLN N N 120.5 0.2 1 43 . 24 ILE H H 8.37 0.02 1 44 . 24 ILE CA C 66.4 0.2 1 45 . 24 ILE N N 119.6 0.2 1 46 . 25 ALA H H 7.90 0.02 1 47 . 25 ALA CA C 55.9 0.2 1 48 . 25 ALA N N 121.3 0.2 1 49 . 26 GLU H H 7.71 0.02 1 50 . 26 GLU CA C 59.8 0.2 1 51 . 26 GLU N N 119.8 0.2 1 52 . 27 PHE H H 8.50 0.02 1 53 . 27 PHE CA C 60.1 0.2 1 54 . 27 PHE N N 120.2 0.2 1 55 . 28 LYS H H 9.13 0.02 1 56 . 28 LYS CA C 60.5 0.2 1 57 . 28 LYS N N 123.5 0.2 1 58 . 29 GLU H H 7.76 0.02 1 59 . 29 GLU CA C 59.7 0.2 1 60 . 29 GLU N N 120.7 0.2 1 61 . 30 ALA H H 7.94 0.02 1 62 . 30 ALA CA C 55.7 0.2 1 63 . 30 ALA N N 122.7 0.2 1 64 . 31 PHE H H 8.74 0.02 1 65 . 31 PHE CA C 62.4 0.2 1 66 . 31 PHE N N 119.3 0.2 1 67 . 32 SER H H 7.85 0.02 1 68 . 32 SER CA C 61.9 0.2 1 69 . 32 SER N N 112.4 0.2 1 70 . 33 LEU H H 7.40 0.02 1 71 . 33 LEU CA C 59.6 0.2 1 72 . 33 LEU N N 121.2 0.2 1 73 . 34 PHE H H 7.33 0.02 1 74 . 34 PHE CA C 59.7 0.2 1 75 . 34 PHE N N 115.0 0.2 1 76 . 35 ASP H H 7.71 0.02 1 77 . 35 ASP CA C 52.8 0.2 1 78 . 35 ASP N N 118.6 0.2 1 79 . 36 LYS H H 7.59 0.02 1 80 . 36 LYS CA C 58.9 0.2 1 81 . 36 LYS N N 124.5 0.2 1 82 . 37 ASP H H 7.98 0.02 1 83 . 37 ASP CA C 53.1 0.2 1 84 . 37 ASP N N 114.0 0.2 1 85 . 38 GLY H H 7.58 0.02 1 86 . 38 GLY CA C 47.6 0.2 1 87 . 38 GLY N N 109.4 0.2 1 88 . 39 ASP H H 8.27 0.02 1 89 . 39 ASP CA C 54.1 0.2 1 90 . 39 ASP N N 120.6 0.2 1 91 . 40 GLY H H 10.52 0.02 1 92 . 40 GLY CA C 45.9 0.2 1 93 . 40 GLY N N 113.3 0.2 1 94 . 41 THR H H 8.02 0.02 1 95 . 41 THR CA C 60.2 0.2 1 96 . 41 THR N N 112.8 0.2 1 97 . 42 ILE H H 9.69 0.02 1 98 . 42 ILE CA C 52.8 0.2 1 99 . 42 ILE N N 127.1 0.2 1 100 . 43 THR H H 8.37 0.02 1 101 . 43 THR CA C 60.0 0.2 1 102 . 43 THR N N 116.6 0.2 1 103 . 44 THR H H 8.98 0.02 1 104 . 44 THR CA C 66.9 0.2 1 105 . 44 THR N N 113.1 0.2 1 106 . 45 LYS H H 8.04 0.02 1 107 . 45 LYS CA C 59.9 0.2 1 108 . 45 LYS N N 119.4 0.2 1 109 . 46 GLU H H 7.56 0.02 1 110 . 46 GLU CA C 59.5 0.2 1 111 . 46 GLU N N 121.3 0.2 1 112 . 47 LEU H H 8.67 0.02 1 113 . 47 LEU CA C 58.3 0.2 1 114 . 47 LEU N N 121.0 0.2 1 115 . 48 GLY H H 8.48 0.02 1 116 . 48 GLY CA C 48.8 0.2 1 117 . 48 GLY N N 105.9 0.2 1 118 . 49 THR H H 7.71 0.02 1 119 . 49 THR CA C 67.1 0.2 1 120 . 49 THR N N 117.9 0.2 1 121 . 50 VAL H H 7.53 0.02 1 122 . 50 VAL CA C 66.5 0.2 1 123 . 50 VAL N N 122.3 0.2 1 124 . 51 MET H H 8.36 0.02 1 125 . 51 MET CA C 59.2 0.2 1 126 . 51 MET N N 118.4 0.2 1 127 . 52 ARG H H 8.37 0.02 1 128 . 52 ARG CA C 59.5 0.2 1 129 . 52 ARG N N 119.3 0.2 1 130 . 53 SER H H 7.82 0.02 1 131 . 53 SER CA C 62.0 0.2 1 132 . 53 SER N N 119.0 0.2 1 133 . 54 LEU H H 7.29 0.02 1 134 . 54 LEU CA C 54.9 0.2 1 135 . 54 LEU N N 121.1 0.2 1 136 . 55 GLY H H 7.78 0.02 1 137 . 55 GLY CA C 45.9 0.2 1 138 . 55 GLY N N 107.0 0.2 1 139 . 56 GLN H H 7.71 0.02 1 140 . 56 GLN CA C 54.8 0.2 1 141 . 56 GLN N N 118.6 0.2 1 142 . 57 ASN H H 8.63 0.02 1 143 . 57 ASN N N 116.5 0.2 1 144 . 58 PRO CA C 62.7 0.2 1 145 . 59 THR H H 8.69 0.02 1 146 . 59 THR CA C 60.9 0.2 1 147 . 59 THR N N 113.2 0.2 1 148 . 60 GLU H H 8.71 0.02 1 149 . 60 GLU CA C 60.5 0.2 1 150 . 60 GLU N N 120.6 0.2 1 151 . 61 ALA H H 8.18 0.02 1 152 . 61 ALA CA C 55.4 0.2 1 153 . 61 ALA N N 120.9 0.2 1 154 . 62 GLU H H 7.59 0.02 1 155 . 62 GLU CA C 59.4 0.2 1 156 . 62 GLU N N 118.7 0.2 1 157 . 63 LEU H H 8.08 0.02 1 158 . 63 LEU CA C 58.2 0.2 1 159 . 63 LEU N N 120.5 0.2 1 160 . 64 GLN H H 8.04 0.02 1 161 . 64 GLN CA C 58.9 0.2 1 162 . 64 GLN N N 118.4 0.2 1 163 . 65 ASP H H 8.00 0.02 1 164 . 65 ASP CA C 57.9 0.2 1 165 . 65 ASP N N 119.9 0.2 1 166 . 66 MET H H 7.75 0.02 1 167 . 66 MET CA C 59.8 0.2 1 168 . 66 MET N N 119.4 0.2 1 169 . 67 ILE H H 7.60 0.02 1 170 . 67 ILE CA C 65.2 0.2 1 171 . 67 ILE N N 117.8 0.2 1 172 . 68 ASN H H 8.42 0.02 1 173 . 68 ASN CA C 56.2 0.2 1 174 . 68 ASN N N 117.5 0.2 1 175 . 69 GLU H H 7.51 0.02 1 176 . 69 GLU CA C 59.4 0.2 1 177 . 69 GLU N N 116.9 0.2 1 178 . 70 VAL H H 7.10 0.02 1 179 . 70 VAL CA C 60.9 0.2 1 180 . 70 VAL N N 107.4 0.2 1 181 . 71 ASP H H 7.60 0.02 1 182 . 71 ASP CA C 54.2 0.2 1 183 . 71 ASP N N 122.2 0.2 1 184 . 72 ALA H H 8.39 0.02 1 185 . 72 ALA CA C 54.7 0.2 1 186 . 72 ALA N N 131.9 0.2 1 187 . 73 ASP H H 8.02 0.02 1 188 . 73 ASP CA C 53.0 0.2 1 189 . 73 ASP N N 113.6 0.2 1 190 . 74 GLY H H 7.46 0.02 1 191 . 74 GLY CA C 47.6 0.2 1 192 . 74 GLY N N 108.3 0.2 1 193 . 75 ASN H H 7.94 0.02 1 194 . 75 ASN CA C 53.0 0.2 1 195 . 75 ASN N N 118.3 0.2 1 196 . 76 GLY H H 10.53 0.02 1 197 . 76 GLY CA C 46.1 0.2 1 198 . 76 GLY N N 113.7 0.2 1 199 . 77 THR H H 7.57 0.02 1 200 . 77 THR CA C 59.9 0.2 1 201 . 77 THR N N 109.0 0.2 1 202 . 78 ILE H H 8.86 0.02 1 203 . 78 ILE CA C 60.0 0.2 1 204 . 78 ILE N N 123.6 0.2 1 205 . 79 ASP H H 8.67 0.02 1 206 . 79 ASP CA C 52.4 0.2 1 207 . 79 ASP N N 128.2 0.2 1 208 . 80 PHE H H 8.75 0.02 1 209 . 80 PHE CA C 64.1 0.2 1 210 . 80 PHE N N 118.5 0.2 1 211 . 81 PRO CA C 66.9 0.2 1 212 . 82 GLU H H 7.79 0.02 1 213 . 82 GLU CA C 59.4 0.2 1 214 . 82 GLU N N 117.2 0.2 1 215 . 83 PHE H H 8.74 0.02 1 216 . 83 PHE CA C 61.5 0.2 1 217 . 83 PHE N N 122.9 0.2 1 218 . 84 LEU H H 8.17 0.02 1 219 . 84 LEU CA C 58.1 0.2 1 220 . 84 LEU N N 118.9 0.2 1 221 . 85 ASN H H 7.12 0.02 1 222 . 85 ASN CA C 56.1 0.2 1 223 . 85 ASN N N 116.3 0.2 1 224 . 86 LEU H H 7.73 0.02 1 225 . 86 LEU CA C 58.5 0.2 1 226 . 86 LEU N N 121.5 0.2 1 227 . 87 MET H H 7.90 0.02 1 228 . 87 MET CA C 56.2 0.2 1 229 . 87 MET N N 116.0 0.2 1 230 . 88 ALA H H 7.92 0.02 1 231 . 88 ALA CA C 54.7 0.2 1 232 . 88 ALA N N 121.7 0.2 1 233 . 89 ARG H H 7.46 0.02 1 234 . 89 ARG CA C 58.3 0.2 1 235 . 89 ARG N N 116.8 0.2 1 236 . 90 LYS H H 7.77 0.02 1 237 . 90 LYS CA C 57.1 0.2 1 238 . 90 LYS N N 118.9 0.2 1 239 . 91 MET H H 7.88 0.02 1 240 . 91 MET CA C 57.0 0.2 1 241 . 91 MET N N 119.1 0.2 1 242 . 92 LYS H H 7.72 0.02 1 243 . 92 LYS CA C 57.0 0.2 1 244 . 92 LYS N N 120.6 0.2 1 245 . 93 ASP H H 8.17 0.02 1 246 . 93 ASP CA C 55.1 0.2 1 247 . 93 ASP N N 121.9 0.2 1 248 . 94 THR H H 8.02 0.02 1 249 . 94 THR CA C 62.8 0.2 1 250 . 94 THR N N 114.8 0.2 1 251 . 95 ASP H H 8.34 0.02 1 252 . 95 ASP CA C 55.3 0.2 1 253 . 95 ASP N N 123.4 0.2 1 254 . 96 SER H H 8.35 0.02 1 255 . 96 SER CA C 60.0 0.2 1 256 . 96 SER N N 117.3 0.2 1 257 . 97 GLU H H 8.36 0.02 1 258 . 97 GLU CA C 58.8 0.2 1 259 . 97 GLU N N 122.5 0.2 1 260 . 98 GLU H H 8.16 0.02 1 261 . 98 GLU CA C 59.7 0.2 1 262 . 98 GLU N N 119.4 0.2 1 263 . 99 GLU H H 8.16 0.02 1 264 . 99 GLU CA C 59.9 0.2 1 265 . 99 GLU N N 119.5 0.2 1 266 . 100 LEU H H 7.98 0.02 1 267 . 100 LEU CA C 58.4 0.2 1 268 . 100 LEU N N 121.7 0.2 1 269 . 101 LYS H H 8.26 0.02 1 270 . 101 LYS CA C 60.6 0.2 1 271 . 101 LYS N N 121.0 0.2 1 272 . 102 GLU H H 7.97 0.02 1 273 . 102 GLU CA C 59.4 0.2 1 274 . 102 GLU N N 119.1 0.2 1 275 . 103 ALA H H 7.97 0.02 1 276 . 103 ALA CA C 55.6 0.2 1 277 . 103 ALA N N 121.9 0.2 1 278 . 104 PHE H H 8.48 0.02 1 279 . 104 PHE CA C 62.5 0.2 1 280 . 104 PHE N N 118.8 0.2 1 281 . 105 ARG H H 7.65 0.02 1 282 . 105 ARG CA C 59.3 0.2 1 283 . 105 ARG N N 116.3 0.2 1 284 . 106 VAL H H 7.34 0.02 1 285 . 106 VAL CA C 66.3 0.2 1 286 . 106 VAL N N 118.6 0.2 1 287 . 107 PHE H H 6.97 0.02 1 288 . 107 PHE CA C 60.5 0.2 1 289 . 107 PHE N N 115.1 0.2 1 290 . 108 ASP H H 7.81 0.02 1 291 . 108 ASP CA C 52.6 0.2 1 292 . 108 ASP N N 116.5 0.2 1 293 . 109 LYS H H 7.70 0.02 1 294 . 109 LYS CA C 59.3 0.2 1 295 . 109 LYS N N 125.7 0.2 1 296 . 110 ASP H H 8.17 0.02 1 297 . 110 ASP CA C 53.4 0.2 1 298 . 110 ASP N N 114.3 0.2 1 299 . 111 GLY H H 7.71 0.02 1 300 . 111 GLY CA C 47.5 0.2 1 301 . 111 GLY N N 109.4 0.2 1 302 . 112 ASN H H 8.30 0.02 1 303 . 112 ASN CA C 53.0 0.2 1 304 . 112 ASN N N 119.8 0.2 1 305 . 113 GLY H H 10.53 0.02 1 306 . 113 GLY CA C 45.3 0.2 1 307 . 113 GLY N N 112.7 0.2 1 308 . 114 PHE H H 7.60 0.02 1 309 . 114 PHE CA C 56.2 0.2 1 310 . 114 PHE N N 116.0 0.2 1 311 . 115 ILE H H 10.18 0.02 1 312 . 115 ILE CA C 61.6 0.2 1 313 . 115 ILE N N 127.6 0.2 1 314 . 116 SER H H 8.90 0.02 1 315 . 116 SER CA C 56.0 0.2 1 316 . 116 SER N N 123.8 0.2 1 317 . 117 ALA H H 9.16 0.02 1 318 . 117 ALA CA C 56.3 0.2 1 319 . 117 ALA N N 123.2 0.2 1 320 . 118 ALA H H 8.16 0.02 1 321 . 118 ALA CA C 55.5 0.2 1 322 . 118 ALA N N 118.4 0.2 1 323 . 119 GLU H H 7.80 0.02 1 324 . 119 GLU CA C 59.8 0.2 1 325 . 119 GLU N N 120.5 0.2 1 326 . 120 LEU H H 8.40 0.02 1 327 . 120 LEU CA C 58.8 0.2 1 328 . 120 LEU N N 121.2 0.2 1 329 . 121 ARG H H 8.54 0.02 1 330 . 121 ARG CA C 60.2 0.2 1 331 . 121 ARG N N 117.7 0.2 1 332 . 122 HIS H H 7.88 0.02 1 333 . 122 HIS CA C 60.5 0.2 1 334 . 122 HIS N N 119.7 0.2 1 335 . 123 VAL H H 7.97 0.02 1 336 . 123 VAL CA C 60.0 0.2 1 337 . 123 VAL N N 119.1 0.2 1 338 . 124 MET H H 8.27 0.02 1 339 . 124 MET CA C 57.7 0.2 1 340 . 124 MET N N 116.3 0.2 1 341 . 125 THR H H 8.15 0.02 1 342 . 125 THR CA C 66.6 0.2 1 343 . 125 THR N N 115.5 0.2 1 344 . 126 ASN H H 7.82 0.02 1 345 . 126 ASN CA C 56.3 0.2 1 346 . 126 ASN N N 122.5 0.2 1 347 . 127 LEU H H 7.74 0.02 1 348 . 127 LEU CA C 55.7 0.2 1 349 . 127 LEU N N 118.9 0.2 1 350 . 128 GLY H H 7.73 0.02 1 351 . 128 GLY CA C 45.7 0.2 1 352 . 128 GLY N N 106.9 0.2 1 353 . 129 GLU H H 7.78 0.02 1 354 . 129 GLU CA C 55.4 0.2 1 355 . 129 GLU N N 120.2 0.2 1 356 . 130 LYS H H 8.53 0.02 1 357 . 130 LYS CA C 55.9 0.2 1 358 . 130 LYS N N 124.5 0.2 1 359 . 131 LEU H H 7.95 0.02 1 360 . 131 LEU CA C 54.5 0.2 1 361 . 131 LEU N N 124.7 0.2 1 362 . 132 THR H H 9.17 0.02 1 363 . 132 THR CA C 61.0 0.2 1 364 . 132 THR N N 114.9 0.2 1 365 . 133 ASP H H 8.81 0.02 1 366 . 133 ASP CA C 58.4 0.2 1 367 . 133 ASP N N 121.2 0.2 1 368 . 134 GLU H H 8.58 0.02 1 369 . 134 GLU CA C 60.4 0.2 1 370 . 134 GLU N N 119.3 0.2 1 371 . 135 GLU H H 7.64 0.02 1 372 . 135 GLU CA C 59.7 0.2 1 373 . 135 GLU N N 120.7 0.2 1 374 . 136 VAL H H 7.90 0.02 1 375 . 136 VAL CA C 67.3 0.2 1 376 . 136 VAL N N 120.9 0.2 1 377 . 137 ASP H H 7.90 0.02 1 378 . 137 ASP CA C 58.0 0.2 1 379 . 137 ASP N N 119.4 0.2 1 380 . 138 GLU H H 7.96 0.02 1 381 . 138 GLU CA C 59.6 0.2 1 382 . 138 GLU N N 119.8 0.2 1 383 . 139 MET H H 7.60 0.02 1 384 . 139 MET CA C 59.6 0.2 1 385 . 139 MET N N 120.6 0.2 1 386 . 140 ILE H H 7.87 0.02 1 387 . 140 ILE CA C 63.9 0.2 1 388 . 140 ILE N N 119.1 0.2 1 389 . 141 ARG H H 8.30 0.02 1 390 . 141 ARG CA C 60.2 0.2 1 391 . 141 ARG N N 118.7 0.2 1 392 . 142 GLU H H 7.93 0.02 1 393 . 142 GLU CA C 59.1 0.2 1 394 . 142 GLU N N 116.5 0.2 1 395 . 143 ALA H H 7.16 0.02 1 396 . 143 ALA CA C 52.1 0.2 1 397 . 143 ALA N N 118.0 0.2 1 398 . 144 ASP H H 8.01 0.02 1 399 . 144 ASP CA C 54.7 0.2 1 400 . 144 ASP N N 118.0 0.2 1 401 . 145 VAL H H 8.27 0.02 1 402 . 145 VAL CA C 65.0 0.2 1 403 . 145 VAL N N 127.3 0.2 1 404 . 146 ASP H H 8.22 0.02 1 405 . 146 ASP CA C 54.1 0.2 1 406 . 146 ASP N N 117.1 0.2 1 407 . 147 GLY H H 7.53 0.02 1 408 . 147 GLY CA C 47.8 0.2 1 409 . 147 GLY N N 108.7 0.2 1 410 . 148 ASP H H 8.30 0.02 1 411 . 148 ASP CA C 54.0 0.2 1 412 . 148 ASP N N 121.0 0.2 1 413 . 149 GLY H H 10.18 0.02 1 414 . 149 GLY CA C 46.2 0.2 1 415 . 149 GLY N N 112.9 0.2 1 416 . 150 GLN H H 7.86 0.02 1 417 . 150 GLN CA C 53.6 0.2 1 418 . 150 GLN N N 115.2 0.2 1 419 . 151 VAL H H 9.06 0.02 1 420 . 151 VAL CA C 62.2 0.2 1 421 . 151 VAL N N 125.7 0.2 1 422 . 152 ASN H H 9.48 0.02 1 423 . 152 ASN CA C 51.6 0.2 1 424 . 152 ASN N N 129.4 0.2 1 425 . 153 TYR H H 8.32 0.02 1 426 . 153 TYR CA C 62.8 0.2 1 427 . 153 TYR N N 118.8 0.2 1 428 . 154 GLU H H 7.93 0.02 1 429 . 154 GLU CA C 60.7 0.2 1 430 . 154 GLU N N 118.7 0.2 1 431 . 155 GLU H H 8.58 0.02 1 432 . 155 GLU CA C 58.8 0.2 1 433 . 155 GLU N N 119.9 0.2 1 434 . 156 PHE H H 8.50 0.02 1 435 . 156 PHE CA C 60.9 0.2 1 436 . 156 PHE N N 123.3 0.2 1 437 . 157 VAL H H 8.33 0.02 1 438 . 157 VAL CA C 67.4 0.2 1 439 . 157 VAL N N 118.5 0.2 1 440 . 158 GLN H H 7.20 0.02 1 441 . 158 GLN CA C 59.1 0.2 1 442 . 158 GLN N N 114.9 0.2 1 443 . 159 VAL H H 7.26 0.02 1 444 . 159 VAL CA C 64.6 0.2 1 445 . 159 VAL N N 115.6 0.2 1 446 . 160 MET H H 7.39 0.02 1 447 . 160 MET CA C 56.0 0.2 1 448 . 160 MET N N 118.9 0.2 1 449 . 161 MET H H 7.60 0.02 1 450 . 161 MET CA C 55.7 0.2 1 451 . 161 MET N N 117.0 0.2 1 452 . 162 ALA H H 7.44 0.02 1 453 . 162 ALA CA C 53.2 0.2 1 454 . 162 ALA N N 124.3 0.2 1 455 . 163 LYS H H 7.66 0.02 1 456 . 163 LYS CA C 57.9 0.2 1 457 . 163 LYS N N 125.8 0.2 1 stop_ save_