data_5184 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H Chemical Shift Assignments for toxin BeKm from the Scorpion Buthus eupeus ; _BMRB_accession_number 5184 _BMRB_flat_file_name bmr5184.str _Entry_type original _Submission_date 2001-10-19 _Accession_date 2001-10-19 _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 Bocharov Eduard V. . 2 Maslennikov Innokenty V. . 3 Korolkova Yuliya V. . 4 Grishin Eugene V. . 5 Arseniev Alexandr S. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 coupling_constants 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 214 "coupling constants" 30 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2002-05-08 original author . stop_ _Original_release_date 2002-05-08 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'Inverse active site of scorpion toxin BeKm' _Citation_status 'in preparation' _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Bocharov Eduard V. . 2 Maslennikov Innokenty V. . 3 Korolkova Yuliya V. . 4 Grishin Eugene V. . 5 Arseniev Alexandr S. . stop_ _Journal_abbreviation 'Nat. Struct. Biol.' _Journal_volume . _Journal_issue . _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first . _Page_last . _Year . _Details . loop_ _Keyword 'scorpion toxin BeKm' 'potassium channel' mutants activity NMR 'spatial structure' stop_ save_ ####################################### # Cited references within the entry # ####################################### save_ref-1 _Saveframe_category citation _Citation_full ; Bartels C., Xia T.-H., Billeter M., Guntert P. & Wuthrich K. The program XEASY for computer-supported NMR spectral analysis of biological macromolecules, J. Biomol. NMR (1995) 6, 1-10. ; _Citation_title . _Citation_status . _Citation_type . _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? _Journal_abbreviation . _Journal_name_full . _Journal_volume . _Journal_issue . _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first . _Page_last . _Year . _Details . save_ save_ref-2 _Saveframe_category citation _Citation_full ; Guntert P., Mumenthaler C. & Wuthrich K. Torsion angle dynamics for NMR structure calculation with new program DYANA, J. Mol. Biol. (1997) 273, 283-298. ; _Citation_title 'Torsion angle dynamics for NMR structure calculation with the new program DYANA.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 9367762 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Guntert P. . . 2 Mumenthaler C. . . 3 Wuthrich K. . . stop_ _Journal_abbreviation 'J. Mol. Biol.' _Journal_name_full 'Journal of molecular biology' _Journal_volume 273 _Journal_issue 1 _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first 283 _Page_last 298 _Year 1997 _Details ; The new program DYANA (DYnamics Algorithm for Nmr Applications) for efficient calculation of three-dimensional protein and nucleic acid structures from distance constraints and torsion angle constraints collected by nuclear magnetic resonance (NMR) experiments performs simulated annealing by molecular dynamics in torsion angle space and uses a fast recursive algorithm to integrate the equations of motions. Torsion angle dynamics can be more efficient than molecular dynamics in Cartesian coordinate space because of the reduced number of degrees of freedom and the concomitant absence of high-frequency bond and angle vibrations, which allows for the use of longer time-steps and/or higher temperatures in the structure calculation. It also represents a significant advance over the variable target function method in torsion angle space with the REDAC strategy used by the predecessor program DIANA. DYANA computation times per accepted conformer in the "bundle" used to represent the NMR structure compare favorably with those of other presently available structure calculation algorithms, and are of the order of 160 seconds for a protein of 165 amino acid residues when using a DEC Alpha 8400 5/300 computer. Test calculations starting from conformers with random torsion angle values further showed that DYANA is capable of efficient calculation of high-quality protein structures with up to 400 amino acid residues, and of nucleic acid structures. ; save_ save_ref-3 _Saveframe_category citation _Citation_full ; Schaumann T., Braun W. & Wuthrich K. (1990) The program FANTOM for energy refinement of polypeptides and proteins using a Newton-Raphson minimizer in torsion angle space, Biopolymers 29, 679-694. ; _Citation_title . _Citation_status . _Citation_type . _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? _Journal_abbreviation . _Journal_name_full . _Journal_volume . _Journal_issue . _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first . _Page_last . _Year . _Details . save_ save_ref-4 _Saveframe_category citation _Citation_full ; Koradi R., Billiter M. & Wutrich, K (1996) MOLMOL: a program for display and analysis of macromolecular structures, J. Mol. Graphics 14, 51-55. ; _Citation_title 'MOLMOL: a program for display and analysis of macromolecular structures.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 8744573 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Koradi R. . . 2 Billeter M. . . 3 Wuthrich K. . . stop_ _Journal_abbreviation . _Journal_name_full 'Journal of molecular graphics' _Journal_volume 14 _Journal_issue 1 _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first 51 _Page_last 32 _Year 1996 _Details ; MOLMOL is a molecular graphics program for display, analysis, and manipulation of three-dimensional structures of biological macromolecules, with special emphasis on nuclear magnetic resonance (NMR) solution structures of proteins and nucleic acids. MOLMOL has a graphical user interface with menus, dialog boxes, and on-line help. The display possibilities include conventional presentation, as well as novel schematic drawings, with the option of combining different presentations in one view of a molecule. Covalent molecular structures can be modified by addition or removal of individual atoms and bonds, and three-dimensional structures can be manipulated by interactive rotation about individual bonds. Special efforts were made to allow for appropriate display and analysis of the sets of typically 20-40 conformers that are conventionally used to represent the result of an NMR structure determination, using functions for superimposing sets of conformers, calculation of root mean square distance (RMSD) values, identification of hydrogen bonds, checking and displaying violations of NMR constraints, and identification and listing of short distances between pairs of hydrogen atoms. ; save_ save_ref-5 _Saveframe_category citation _Citation_full ; Korolkova YV, Kozlov SA, Lipkin AV, Pluzhnikov KA, Hadley JK, Filippov AK, Brown DA, Angelo K, Strobaek D, Jespersen T, Olesen SP, Jensen BS, Grishin EV. An ERG channel inhibitor from the scorpion Buthus eupeus. J. Biol. Chem. 2001 Mar 30;276(13):9868-76 ; _Citation_title 'An ERG channel inhibitor from the scorpion Buthus eupeus.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11136720 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Korolkova 'Y. V.' V. . 2 Kozlov 'S. A.' A. . 3 Lipkin 'A. V.' V. . 4 Pluzhnikov 'K. A.' A. . 5 Hadley 'J. K.' K. . 6 Filippov 'A. K.' K. . 7 Brown 'D. A.' A. . 8 Angelo K. . . 9 Strobaek D. . . 10 Jespersen T. . . 11 Olesen 'S. P.' P. . 12 Jensen 'B. S.' S. . 13 Grishin 'E. V.' V. . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'The Journal of biological chemistry' _Journal_volume 276 _Journal_issue 13 _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 9868 _Page_last 9876 _Year 2001 _Details ; The isolation of the peptide inhibitor of M-type K(+) current, BeKm-1, from the venom of the Central Asian scorpion Buthus eupeus has been described previously (Fillipov A. K., Kozlov, S. A., Pluzhnikov, K. A., Grishin, E. V., and Brown, D. A. (1996) FEBS Lett. 384, 277-280). Here we report the cloning, expression, and selectivity of BeKm-1. A full-length cDNA of 365 nucleotides encoding the precursor of BeKm-1 was isolated using the rapid amplification of cDNA ends polymerase chain reaction technique from mRNA obtained from scorpion telsons. Sequence analysis of the cDNA revealed that the precursor contains a signal peptide of 21 amino acid residues. The mature toxin consists of 36 amino acid residues. BeKm-1 belongs to the family of scorpion venom potassium channel blockers and represents a new subgroup of these toxins. The recombinant BeKm-1 was produced as a Protein A fusion product in the periplasm of Escherichia coli. After cleavage and high performance liquid chromatography purification, recombinant BeKm-1 displayed the same properties as the native toxin. Three BeKm-1 mutants (R27K, F32K, and R27K/F32K) were generated, purified, and characterized. Recombinant wild-type BeKm-1 and the three mutants partly inhibited the native M-like current in NG108-15 at 100 nm. The effect of the recombinant BeKm-1 on different K(+) channels was also studied. BeKm-1 inhibited hERG1 channels with an IC(50) of 3.3 nm, but had no effect at 100 nm on hEAG, hSK1, rSK2, hIK, hBK, KCNQ1/KCNE1, KCNQ2/KCNQ3, KCNQ4 channels, and minimal effect on rELK1. Thus, BeKm-1 was shown to be a novel specific blocker of hERG1 potassium channels. ; save_ ################################## # Molecular system description # ################################## save_system_BeKm _Saveframe_category molecular_system _Mol_system_name 'toxin BeKm from the scorpion Buthus eupeus' _Abbreviation_common BeKm _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label BeKm $BeKm stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all disulfide bound' loop_ _Biological_function 'Scorpion toxin BeKm is specific blocker of hERG1 potassium channels.' stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_BeKm _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'scorpion toxin BeKm' _Abbreviation_common BeKm _Molecular_mass 4092 _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 36 _Mol_residue_sequence ; RPTDIKCSESYQCFPVCKSR FGKTNGRCVNGFCDCF ; loop_ _Residue_seq_code _Residue_label 1 ARG 2 PRO 3 THR 4 ASP 5 ILE 6 LYS 7 CYS 8 SER 9 GLU 10 SER 11 TYR 12 GLN 13 CYS 14 PHE 15 PRO 16 VAL 17 CYS 18 LYS 19 SER 20 ARG 21 PHE 22 GLY 23 LYS 24 THR 25 ASN 26 GLY 27 ARG 28 CYS 29 VAL 30 ASN 31 GLY 32 PHE 33 CYS 34 ASP 35 CYS 36 PHE 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 PDB 1J5J "Solution Structure Of Herg-Specific Scorpion Toxin Bekm-1" 100.00 36 100.00 100.00 1.71e-16 PDB 1LGL "Solution Structure Of Herg-Specific Scorpion Toxin Bekm-1" 100.00 36 100.00 100.00 1.71e-16 GB AAK28021 "BeKm-1 toxin precursor [Mesobuthus eupeus]" 100.00 57 100.00 100.00 3.71e-17 GB AIL48783 "potassium channel blocker pMeKTx12-1 [Mesobuthus eupeus]" 100.00 57 97.22 97.22 2.26e-16 SP Q9BKB7 "RecName: Full=Potassium channel toxin gamma-KTx 2.1; AltName: Full=Neurotoxin BeKm-1; Flags: Precursor [Mesobuthus eupeus]" 100.00 57 100.00 100.00 3.71e-17 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Secretion $BeKm 'lesser Asian scorpion' 34648 Eukaryota Metazoa Buthus eupeus venom stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Cell_line _Vector_name $BeKm 'recombinant technology' 'E. coli' Escherichia coli . HB101 . 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 $BeKm 1.0 mM . stop_ save_ ############################ # Computer software used # ############################ save_XEASY _Saveframe_category software _Name XEASY _Version 1.2.11 loop_ _Task 'peak assignments' stop_ _Details . _Citation_label $ref-1 save_ save_DYANA _Saveframe_category software _Name DYANA _Version 1.5 loop_ _Task 'structure calculation' stop_ _Details . _Citation_label $ref-2 save_ save_FANTOM _Saveframe_category software _Name FANTOM _Version 4 loop_ _Task 'energy minimization' stop_ _Details . _Citation_label $ref-3 save_ save_MOLMOL _Saveframe_category software _Name MOLMOL _Version 2.5.1 loop_ _Task visualization 'structure analysis' stop_ _Details . _Citation_label $ref-4 save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model UNITY _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_1H_COSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H COSY' _Sample_label $sample_1 save_ save_1H_TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1H TOCSY' _Sample_label $sample_1 save_ save_1H_NOESY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '1H NOESY' _Sample_label $sample_1 save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H COSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1H TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name '1H NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_sample_cond_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 3.5 0.2 n/a temperature 303 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 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_chem_shift_set_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name BeKm _Text_data_format . _Text_data . loop_ _Atom_shift_assign_ID _Residue_author_seq_code _Residue_seq_code _Residue_label _Atom_name _Atom_type _Chem_shift_value _Chem_shift_value_error _Chem_shift_ambiguity_code 1 . 1 ARG HA H 4.37 0.02 1 2 . 1 ARG HB2 H 1.90 0.02 1 3 . 1 ARG HB3 H 1.90 0.02 1 4 . 1 ARG HG2 H 1.64 0.02 1 5 . 1 ARG HG3 H 1.64 0.02 1 6 . 1 ARG HD2 H 3.06 0.02 1 7 . 1 ARG HD3 H 3.06 0.02 1 8 . 1 ARG HE H 7.09 0.02 1 9 . 2 PRO HA H 4.68 0.02 1 10 . 2 PRO HB2 H 1.90 0.02 1 11 . 2 PRO HB3 H 2.33 0.02 1 12 . 2 PRO HG2 H 1.98 0.02 1 13 . 2 PRO HG3 H 1.98 0.02 1 14 . 2 PRO HD2 H 3.58 0.02 1 15 . 2 PRO HD3 H 3.73 0.02 1 16 . 3 THR H H 7.72 0.02 1 17 . 3 THR HA H 4.69 0.02 1 18 . 3 THR HB H 4.26 0.02 1 19 . 3 THR HG2 H 0.87 0.02 1 20 . 4 ASP H H 8.33 0.02 1 21 . 4 ASP HA H 4.68 0.02 1 22 . 4 ASP HB2 H 2.99 0.02 1 23 . 4 ASP HB3 H 2.70 0.02 1 24 . 5 ILE H H 8.24 0.02 1 25 . 5 ILE HA H 4.12 0.02 1 26 . 5 ILE HB H 1.75 0.02 1 27 . 5 ILE HG2 H 0.93 0.02 1 28 . 5 ILE HG12 H 1.33 0.02 1 29 . 5 ILE HG13 H 1.65 0.02 1 30 . 5 ILE HD1 H 0.96 0.02 1 31 . 6 LYS H H 8.34 0.02 1 32 . 6 LYS HA H 4.41 0.02 1 33 . 6 LYS HB2 H 1.57 0.02 1 34 . 6 LYS HB3 H 1.57 0.02 1 35 . 6 LYS HG2 H 1.10 0.02 1 36 . 6 LYS HG3 H 1.10 0.02 1 37 . 6 LYS HD2 H 1.43 0.02 1 38 . 6 LYS HD3 H 1.43 0.02 1 39 . 7 CYS H H 8.00 0.02 1 40 . 7 CYS HA H 4.60 0.02 1 41 . 7 CYS HB2 H 2.95 0.02 1 42 . 7 CYS HB3 H 2.80 0.02 1 43 . 8 SER H H 9.29 0.02 1 44 . 8 SER HA H 4.54 0.02 1 45 . 8 SER HB2 H 3.75 0.02 1 46 . 8 SER HB3 H 3.87 0.02 1 47 . 9 GLU H H 7.66 0.02 1 48 . 9 GLU HA H 4.76 0.02 1 49 . 9 GLU HB2 H 1.31 0.02 1 50 . 9 GLU HB3 H 2.14 0.02 1 51 . 9 GLU HG2 H 2.41 0.02 1 52 . 9 GLU HG3 H 2.43 0.02 1 53 . 10 SER H H 9.33 0.02 1 54 . 10 SER HA H 3.91 0.02 1 55 . 10 SER HB2 H 4.18 0.02 1 56 . 10 SER HB3 H 4.02 0.02 1 57 . 11 TYR H H 8.35 0.02 1 58 . 11 TYR HA H 3.70 0.02 1 59 . 11 TYR HB2 H 3.13 0.02 1 60 . 11 TYR HB3 H 2.94 0.02 1 61 . 11 TYR HD1 H 7.06 0.02 1 62 . 11 TYR HD2 H 7.06 0.02 1 63 . 11 TYR HE1 H 6.81 0.02 1 64 . 11 TYR HE2 H 6.81 0.02 1 65 . 12 GLN H H 6.79 0.02 1 66 . 12 GLN HA H 3.93 0.02 1 67 . 12 GLN HB2 H 2.21 0.02 1 68 . 12 GLN HB3 H 2.01 0.02 1 69 . 12 GLN HG2 H 2.29 0.02 1 70 . 12 GLN HG3 H 1.47 0.02 1 71 . 12 GLN HE21 H 7.30 0.02 1 72 . 12 GLN HE22 H 6.83 0.02 1 73 . 13 CYS H H 7.84 0.02 1 74 . 13 CYS HA H 4.58 0.02 1 75 . 13 CYS HB2 H 2.61 0.02 1 76 . 13 CYS HB3 H 2.95 0.02 1 77 . 14 PHE H H 7.28 0.02 1 78 . 14 PHE HA H 4.32 0.02 1 79 . 14 PHE HB2 H 3.29 0.02 1 80 . 14 PHE HB3 H 3.36 0.02 1 81 . 14 PHE HD1 H 7.26 0.02 1 82 . 14 PHE HD2 H 7.26 0.02 1 83 . 14 PHE HE1 H 7.52 0.02 1 84 . 14 PHE HE2 H 7.52 0.02 1 85 . 14 PHE HZ H 7.42 0.02 1 86 . 15 PRO HA H 4.39 0.02 1 87 . 15 PRO HB2 H 1.99 0.02 1 88 . 15 PRO HB3 H 2.38 0.02 1 89 . 15 PRO HG2 H 2.13 0.02 1 90 . 15 PRO HG3 H 2.13 0.02 1 91 . 15 PRO HD2 H 3.86 0.02 1 92 . 15 PRO HD3 H 3.86 0.02 1 93 . 16 VAL H H 6.63 0.02 1 94 . 16 VAL HA H 4.11 0.02 1 95 . 16 VAL HB H 2.17 0.02 1 96 . 16 VAL HG1 H 1.09 0.02 1 97 . 16 VAL HG2 H 1.14 0.02 1 98 . 17 CYS H H 8.46 0.02 1 99 . 17 CYS HA H 4.76 0.02 1 100 . 17 CYS HB2 H 2.75 0.02 1 101 . 17 CYS HB3 H 2.86 0.02 1 102 . 18 LYS H H 7.66 0.02 1 103 . 18 LYS HA H 4.24 0.02 1 104 . 18 LYS HB2 H 1.83 0.02 1 105 . 18 LYS HB3 H 1.83 0.02 1 106 . 18 LYS HG2 H 1.28 0.02 1 107 . 18 LYS HG3 H 1.35 0.02 1 108 . 18 LYS HD2 H 1.53 0.02 1 109 . 18 LYS HD3 H 1.53 0.02 1 110 . 19 SER H H 8.35 0.02 1 111 . 19 SER HA H 4.18 0.02 1 112 . 19 SER HB2 H 3.98 0.02 1 113 . 19 SER HB3 H 3.98 0.02 1 114 . 20 ARG H H 8.44 0.02 1 115 . 20 ARG HA H 4.19 0.02 1 116 . 20 ARG HB2 H 1.15 0.02 1 117 . 20 ARG HB3 H 1.53 0.02 1 118 . 20 ARG HG2 H 1.38 0.02 1 119 . 20 ARG HG3 H 1.05 0.02 1 120 . 20 ARG HD2 H 2.92 0.02 1 121 . 20 ARG HD3 H 2.92 0.02 1 122 . 20 ARG HE H 7.08 0.02 1 123 . 21 PHE H H 6.94 0.02 1 124 . 21 PHE HA H 4.92 0.02 1 125 . 21 PHE HB2 H 3.06 0.02 1 126 . 21 PHE HB3 H 3.44 0.02 1 127 . 21 PHE HD1 H 7.40 0.02 1 128 . 21 PHE HD2 H 7.40 0.02 1 129 . 21 PHE HE1 H 7.26 0.02 1 130 . 21 PHE HE2 H 7.26 0.02 1 131 . 21 PHE HZ H 7.35 0.02 1 132 . 22 GLY H H 7.69 0.02 1 133 . 22 GLY HA2 H 4.02 0.02 1 134 . 22 GLY HA3 H 4.02 0.02 1 135 . 23 LYS H H 7.79 0.02 1 136 . 23 LYS HA H 4.72 0.02 1 137 . 23 LYS HB2 H 1.38 0.02 1 138 . 23 LYS HB3 H 2.09 0.02 1 139 . 23 LYS HG2 H 1.13 0.02 1 140 . 23 LYS HG3 H 1.41 0.02 1 141 . 23 LYS HD2 H 1.64 0.02 1 142 . 23 LYS HD3 H 1.64 0.02 1 143 . 23 LYS HE2 H 2.98 0.02 1 144 . 23 LYS HE3 H 2.98 0.02 1 145 . 24 THR H H 8.46 0.02 1 146 . 24 THR HA H 4.05 0.02 1 147 . 24 THR HB H 4.30 0.02 1 148 . 24 THR HG2 H 1.13 0.02 1 149 . 25 ASN H H 7.80 0.02 1 150 . 25 ASN HA H 4.67 0.02 1 151 . 25 ASN HB2 H 1.64 0.02 1 152 . 25 ASN HB3 H 1.68 0.02 1 153 . 25 ASN HD21 H 7.36 0.02 1 154 . 25 ASN HD22 H 6.81 0.02 1 155 . 26 GLY H H 7.08 0.02 1 156 . 26 GLY HA2 H 5.01 0.02 1 157 . 26 GLY HA3 H 3.31 0.02 1 158 . 27 ARG H H 8.77 0.02 1 159 . 27 ARG HA H 4.64 0.02 1 160 . 27 ARG HB2 H 1.86 0.02 1 161 . 27 ARG HB3 H 1.86 0.02 1 162 . 27 ARG HG2 H 1.62 0.02 1 163 . 27 ARG HG3 H 1.66 0.02 1 164 . 27 ARG HD2 H 3.14 0.02 1 165 . 27 ARG HD3 H 3.14 0.02 1 166 . 27 ARG HE H 7.18 0.02 1 167 . 28 CYS H H 8.89 0.02 1 168 . 28 CYS HA H 4.80 0.02 1 169 . 28 CYS HB2 H 2.26 0.02 1 170 . 28 CYS HB3 H 2.78 0.02 1 171 . 29 VAL H H 9.07 0.02 1 172 . 29 VAL HA H 4.24 0.02 1 173 . 29 VAL HB H 2.03 0.02 1 174 . 29 VAL HG1 H 0.91 0.02 1 175 . 29 VAL HG2 H 0.87 0.02 1 176 . 30 ASN H H 9.35 0.02 1 177 . 30 ASN HA H 4.33 0.02 1 178 . 30 ASN HB2 H 2.87 0.02 1 179 . 30 ASN HB3 H 3.16 0.02 1 180 . 30 ASN HD21 H 7.65 0.02 1 181 . 30 ASN HD22 H 6.89 0.02 1 182 . 31 GLY H H 7.94 0.02 1 183 . 31 GLY HA2 H 3.75 0.02 1 184 . 31 GLY HA3 H 3.86 0.02 1 185 . 32 PHE H H 7.67 0.02 1 186 . 32 PHE HA H 5.45 0.02 1 187 . 32 PHE HB2 H 2.68 0.02 1 188 . 32 PHE HB3 H 3.20 0.02 1 189 . 32 PHE HD1 H 7.06 0.02 1 190 . 32 PHE HD2 H 7.06 0.02 1 191 . 32 PHE HE1 H 7.27 0.02 1 192 . 32 PHE HE2 H 7.27 0.02 1 193 . 32 PHE HZ H 7.25 0.02 1 194 . 33 CYS H H 9.03 0.02 1 195 . 33 CYS HA H 4.92 0.02 1 196 . 33 CYS HB2 H 2.76 0.02 1 197 . 33 CYS HB3 H 2.56 0.02 1 198 . 34 ASP H H 9.65 0.02 1 199 . 34 ASP HA H 5.13 0.02 1 200 . 34 ASP HB2 H 2.56 0.02 1 201 . 34 ASP HB3 H 2.85 0.02 1 202 . 35 CYS H H 8.06 0.02 1 203 . 35 CYS HA H 5.49 0.02 1 204 . 35 CYS HB2 H 2.35 0.02 1 205 . 35 CYS HB3 H 2.86 0.02 1 206 . 36 PHE H H 8.25 0.02 1 207 . 36 PHE HA H 4.53 0.02 1 208 . 36 PHE HB2 H 3.06 0.02 1 209 . 36 PHE HB3 H 2.94 0.02 1 210 . 36 PHE HD1 H 7.00 0.02 1 211 . 36 PHE HD2 H 7.00 0.02 1 212 . 36 PHE HE1 H 7.02 0.02 1 213 . 36 PHE HE2 H 7.02 0.02 1 214 . 36 PHE HZ H 7.09 0.02 1 stop_ save_ ######################## # Coupling constants # ######################## save_coupling_constants_set_1 _Saveframe_category coupling_constants _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_cond_1 _Spectrometer_frequency_1H 600 _Mol_system_component_name BeKm _Text_data_format . _Text_data . loop_ _Coupling_constant_ID _Coupling_constant_code _Atom_one_residue_seq_code _Atom_one_residue_label _Atom_one_name _Atom_two_residue_seq_code _Atom_two_residue_label _Atom_two_name _Coupling_constant_value _Coupling_constant_min_value _Coupling_constant_max_value _Coupling_constant_value_error 1 3JHNHA 3 THR H 3 THR HA 9.5 . . 1.0 2 3JHNHA 4 ASP H 4 ASP HA 8.5 . . 1.0 3 3JHNHA 5 ILE H 5 ILE HA 7.5 . . 1.0 4 3JHNHA 6 LYS H 6 LYS HA 5.0 . . 1.0 5 3JHNHA 7 CYS H 7 CYS HA 8.0 . . 1.0 6 3JHNHA 8 SER H 8 SER HA 8.3 . . 1.0 7 3JHNHA 9 GLU H 9 GLU HA 5.5 . . 1.0 8 3JHNHA 10 SER H 10 SER HA 5.0 . . 1.0 9 3JHNHA 11 TYR H 11 TYR HA 4.0 . . 1.0 10 3JHNHA 12 GLN H 12 GLN HA 6.0 . . 1.0 11 3JHNHA 13 CYS H 13 CYS HA 6.5 . . 1.0 12 3JHNHA 14 PHE H 14 PHE HA 4.0 . . 1.0 13 3JHNHA 16 VAL H 16 VAL HA 5.0 . . 1.0 14 3JHNHA 17 CYS H 17 CYS HA 4.5 . . 1.0 15 3JHNHA 18 LYS H 18 LYS HA 4.0 . . 1.0 16 3JHNHA 19 SER H 19 SER HA 5.5 . . 1.0 17 3JHNHA 20 ARG H 20 ARG HA 7.5 . . 1.0 18 3JHNHA 21 PHE H 21 PHE HA 7.5 . . 1.0 19 3JHNHA 23 LYS H 23 LYS HA 8.0 . . 1.0 20 3JHNHA 24 THR H 24 THR HA 8.5 . . 1.0 21 3JHNHA 25 ASN H 25 ASN HA 7.5 . . 1.0 22 3JHNHA 27 ARG H 27 ARG HA 8.8 . . 1.0 23 3JHNHA 28 CYS H 28 CYS HA 8.2 . . 1.0 24 3JHNHA 29 VAL H 29 VAL HA 10.4 . . 1.0 25 3JHNHA 30 ASN H 30 ASN HA 6.5 . . 1.0 26 3JHNHA 32 PHE H 32 PHE HA 10.5 . . 1.0 27 3JHNHA 33 CYS H 33 CYS HA 4.5 . . 1.0 28 3JHNHA 34 ASP H 34 ASP HA 9.0 . . 1.0 29 3JHNHA 35 CYS H 35 CYS HA 10.0 . . 1.0 30 3JHNHA 36 PHE H 36 PHE HA 7.0 . . 1.0 stop_ save_