data_6427 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Solution structure of a recombinant type I sculpin antifreeze protein ; _BMRB_accession_number 6427 _BMRB_flat_file_name bmr6427.str _Entry_type original _Submission_date 2004-12-10 _Accession_date 2004-12-10 _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 Kwan A. H.Y. . 2 Fairley K. . . 3 Anderberg P. I. . 4 Liew C. W. . 5 Harding M. M. . 6 Mackay J. P. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 2 residual_dipolar_couplings 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 282 "13C chemical shifts" 204 "15N chemical shifts" 66 "residual dipolar couplings" 18 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2005-06-02 original author . stop_ _Original_release_date 2005-06-02 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'Solution structure of a recombinant type I sculpin antifreeze protein.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 15697223 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Kwan A. H. . 2 Fairley K. . . 3 Anderberg P. I. . 4 Liew C. W. . 5 Harding M. M. . 6 Mackay J. P. . stop_ _Journal_abbreviation Biochemistry _Journal_volume 44 _Journal_issue 6 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 1980 _Page_last 1988 _Year 2005 _Details . loop_ _Keyword a-helix 'NMR spectroscopy' sculpin 'solution structure' 'type I antifreeze protein' stop_ save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Fairley K, Westman BJ, Pham LH, Haymet AD, Harding MM, Mackay JP. Type I shorthorn sculpin antifreeze protein: recombinant synthesis, solution conformation, and ice growth inhibition studies. J Biol Chem. 2002 Jul 5;277(27):24073-80. Epub 2002 Apr 08. ; _Citation_title 'Type I shorthorn sculpin antifreeze protein: recombinant synthesis, solution conformation, and ice growth inhibition studies.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11940576 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Fairley Kayesh . . 2 Westman 'Belinda J' J. . 3 Pham 'Linda H' H. . 4 Haymet 'A D J' D. . 5 Harding 'Margaret M' M. . 6 Mackay 'Joel P' P. . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'The Journal of biological chemistry' _Journal_volume 277 _Journal_issue 27 _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 24073 _Page_last 24080 _Year 2002 _Details ; A number of structurally diverse classes of "antifreeze" proteins that allow fish to survive in sub-zero ice-laden waters have been isolated from the blood plasma of cold water teleosts. However, despite receiving a great deal of attention, the one or more mechanisms through which these proteins act are not fully understood. In this report we have synthesized a type I antifreeze polypeptide (AFP) from the shorthorn sculpin Myoxocephalus scorpius using recombinant methods. Construction of a synthetic gene with optimized codon usage and expression as a glutathione S-transferase fusion protein followed by purification yielded milligram amounts of polypeptide with two extra residues appended to the N terminus. Circular dichroism and NMR experiments, including residual dipolar coupling measurements on a 15N-labeled recombinant polypeptide, show that the polypeptides are alpha-helical with the first four residues being more flexible than the remainder of the sequence. Both the recombinant and synthetic polypeptides modify ice growth, forming facetted crystals just below the freezing point, but display negligible thermal hysteresis. Acetylation of Lys-10, Lys-20, and Lys-21 as well as the N terminus of the recombinant polypeptide gave a derivative that displays both thermal hysteresis (0.4 degrees C at 15 mg/ml) and ice crystal faceting. These results confirm that the N terminus of wild-type polypeptide is functionally important and support our previously proposed mechanism for all type I proteins, in which the hydrophobic face is oriented toward the ice at the ice/water interface. ; save_ ################################## # Molecular system description # ################################## save_system_SS3 _Saveframe_category molecular_system _Mol_system_name 'Antifreeze peptide SS-3' _Abbreviation_common SS3 _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label rSS3 $rSS3 stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'not present' loop_ _Biological_function 'antifreeze protein' stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_rSS3 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common SS3 _Abbreviation_common SS3 _Molecular_mass 3083.5 _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 35 _Mol_residue_sequence ; GSMNAPARAAAKTAADALAA AKKTAADAAAAAAAA ; loop_ _Residue_seq_code _Residue_label 1 GLY 2 SER 3 MET 4 ASN 5 ALA 6 PRO 7 ALA 8 ARG 9 ALA 10 ALA 11 ALA 12 LYS 13 THR 14 ALA 15 ALA 16 ASP 17 ALA 18 LEU 19 ALA 20 ALA 21 ALA 22 LYS 23 LYS 24 THR 25 ALA 26 ALA 27 ASP 28 ALA 29 ALA 30 ALA 31 ALA 32 ALA 33 ALA 34 ALA 35 ALA stop_ _Sequence_homology_query_date 2008-08-19 _Sequence_homology_query_revised_last_date 2008-08-19 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 1Y03 'Solution Structure Of A Recombinant Type I Sculpin Antifreeze Protein' 100.00 35 100.00 100.00 2.12e-05 PDB 1Y04 'Solution Structure Of A Recombinant Type I Sculpin Antifreeze Protein' 100.00 35 100.00 100.00 2.12e-05 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $rSS3 'Shorthorn sculpin' 8097 Eukaryota Metazoa Myoxocephalus scorpius 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 $rSS3 'recombinant technology' . . . . . 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 _Concentration_min_value _Concentration_max_value _Isotopic_labeling $rSS3 . mM 0.8 1.2 . H2O 95 % . . . D2O 5 % . . . stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $rSS3 . mM 0.8 1.2 '[U-15N; U-13C]' H2O 95 % . . . D2O 5 % . . . stop_ save_ save_sample_3 _Saveframe_category sample _Sample_type Bi-cell _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $rSS3 . mM 0.1 0.3 [U-15N] stop_ save_ ############################ # Computer software used # ############################ save_XWINNMR _Saveframe_category software _Name xwinnmr _Version 2.5 loop_ _Task processing stop_ _Details Bruker save_ save_XEASY _Saveframe_category software _Name XEASY _Version 1.3.13 loop_ _Task 'data analysis' stop_ _Details ; Bartels, C.; Xia, T.-H.; Billeter, P.; Guntert, P.; Wuthrich, K. J. Biomol. NMR 1995, 5, 1-10. ; save_ save_SPARKY _Saveframe_category software _Name SPARKY _Version 3 loop_ _Task 'data analysis' stop_ _Details ; Goddard, T.D. and Kneller, D.G. SAPRKY 3, University of California, San Francisco, USA. ; save_ save_ARIA _Saveframe_category software _Name ARIA _Version 1.2 loop_ _Task 'structure solution, refinement' stop_ _Details ; Nilges, M. Fold Des 1997, 2, S53-57. Linge, J. P.; O'Donoghue, S. I.; Nilges, M. Meth. Enzymol. 2001, 339, 71-90. Nilges, M.; Macias, M. J.; O'Donoghue, S. I.; Oschkinat, H. J. Mol. Biol. 1997, 269, 408-422. Nilges, M. J. Mol. Biol. 1995, 245, 645-660. ; save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DRX _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_TOCSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D TOCSY' _Sample_label . save_ save_2D_NOESY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D NOESY' _Sample_label . save_ save_DQF-COSY_3 _Saveframe_category NMR_applied_experiment _Experiment_name DQF-COSY _Sample_label . save_ save_HNHA_4 _Saveframe_category NMR_applied_experiment _Experiment_name HNHA _Sample_label . save_ save_HNCA_5 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA _Sample_label . save_ save_HNCACB_6 _Saveframe_category NMR_applied_experiment _Experiment_name HNCACB _Sample_label . save_ save_CBCA(CO)NH_7 _Saveframe_category NMR_applied_experiment _Experiment_name CBCA(CO)NH _Sample_label . save_ save_HNCO_8 _Saveframe_category NMR_applied_experiment _Experiment_name HNCO _Sample_label . save_ save_HNCA-ECOSY_9 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA-ECOSY _Sample_label . save_ save_HET-NOE_10 _Saveframe_category NMR_applied_experiment _Experiment_name HET-NOE _Sample_label . save_ save_15N-HSQC_11 _Saveframe_category NMR_applied_experiment _Experiment_name 15N-HSQC _Sample_label . save_ save_13C-HSQC_12 _Saveframe_category NMR_applied_experiment _Experiment_name 13C-HSQC _Sample_label . save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name DQF-COSY _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_4 _Saveframe_category NMR_applied_experiment _Experiment_name HNHA _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_5 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_6 _Saveframe_category NMR_applied_experiment _Experiment_name HNCACB _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_7 _Saveframe_category NMR_applied_experiment _Experiment_name CBCA(CO)NH _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_8 _Saveframe_category NMR_applied_experiment _Experiment_name HNCO _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_9 _Saveframe_category NMR_applied_experiment _Experiment_name HNCA-ECOSY _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_10 _Saveframe_category NMR_applied_experiment _Experiment_name HET-NOE _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_11 _Saveframe_category NMR_applied_experiment _Experiment_name 15N-HSQC _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_12 _Saveframe_category NMR_applied_experiment _Experiment_name 13C-HSQC _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_sample_cond_1 _Saveframe_category sample_conditions _Details 'pH is adjusted to 5.0 using NaOH.' loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' . . mM pH 5.0 0.2 pH pressure 1 . atm temperature 268 1 K stop_ save_ save_sample_cond_2 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' . . mM pH 5.0 0.2 pH pressure 1 . atm temperature 278 1 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 C 13 'methyl protons' ppm 0.0 . indirect . . . 0.251449530 DSS H 1 'methyl protons' ppm 0.0 internal direct . . . 1.0 DSS N 15 'methyl protons' ppm 0.0 . indirect . . . 0.101329118 stop_ save_ ################################### # Assigned chemical shift lists # ################################### ################################################################### # Chemical Shift Ambiguity Index Value Definitions # # # # The values other than 1 are used for those atoms with different # # chemical shifts that cannot be assigned to stereospecific atoms # # or to specific residues or chains. # # # # Index Value Definition # # # # 1 Unique (including isolated methyl protons, # # geminal atoms, and geminal methyl # # groups with identical chemical shifts) # # (e.g. ILE HD11, HD12, HD13 protons) # # 2 Ambiguity of geminal atoms or geminal methyl # # proton groups (e.g. ASP HB2 and HB3 # # protons, LEU CD1 and CD2 carbons, or # # LEU HD11, HD12, HD13 and HD21, HD22, # # HD23 methyl protons) # # 3 Aromatic atoms on opposite sides of # # symmetrical rings (e.g. TYR HE1 and HE2 # # protons) # # 4 Intraresidue ambiguities (e.g. LYS HG and # # HD protons or TRP HZ2 and HZ3 protons) # # 5 Interresidue ambiguities (LYS 12 vs. LYS 27) # # 6 Intermolecular ambiguities (e.g. ASP 31 CA # # in monomer 1 and ASP 31 CA in monomer 2 # # of an asymmetrical homodimer, duplex # # DNA assignments, or other assignments # # that may apply to atoms in one or more # # molecule in the molecular assembly) # # 9 Ambiguous, specific ambiguity not defined # # # ################################################################### save_chemical_shift_set_268K _Saveframe_category assigned_chemical_shifts _Details . loop_ _Sample_label $sample_1 $sample_2 stop_ _Sample_conditions_label $sample_cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name rSS3 _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 GLY CA C 42.98 0.2 . 2 . 1 GLY C C 170.07 0.2 . 3 . 2 SER H H 8.73 0.02 . 4 . 2 SER N N 115.63 0.2 . 5 . 2 SER CA C 58.14 0.2 . 6 . 2 SER HA H 4.5 0.02 . 7 . 2 SER C C 174.6 0.2 . 8 . 2 SER CB C 63.6 0.2 . 9 . 2 SER HB2 H 3.85 0.02 . 10 . 2 SER HB3 H 3.89 0.02 . 11 . 3 MET H H 8.67 0.02 . 12 . 3 MET N N 122.69 0.2 . 13 . 3 MET CA C 55.45 0.04 . 14 . 3 MET HA H 4.52 0.02 . 15 . 3 MET C C 175.85 0.2 . 16 . 3 MET CB C 32.6 0.05 . 17 . 3 MET HB2 H 2.02 0.02 . 18 . 3 MET HB3 H 2.08 0.02 . 19 . 3 MET HG2 H 2.55 0.02 . 20 . 3 MET HG3 H 2.61 0.02 . 21 . 4 ASN H H 8.44 0.02 . 22 . 4 ASN N N 120.37 0.2 . 23 . 4 ASN CA C 52.41 0.2 . 24 . 4 ASN HA H 4.71 0.02 . 25 . 4 ASN C C 174.48 0.2 . 26 . 4 ASN CB C 38.4 0.01 . 27 . 4 ASN HB2 H 1.45 0.02 . 28 . 4 ASN HB3 H 2.87 0.02 . 29 . 4 ASN HD21 H 7.05 0.02 . 30 . 4 ASN HD22 H 7.8 0.02 . 31 . 5 ALA H H 8.45 0.02 . 32 . 5 ALA N N 124.91 0.2 . 33 . 5 ALA CA C 51.97 0.2 . 34 . 5 ALA CB C 18.94 0.2 . 35 . 5 ALA HB H 1.44 0.02 . 36 . 6 PRO CA C 64.04 0.2 . 37 . 6 PRO HA H 4.42 0.02 . 38 . 6 PRO C C 177.94 0.2 . 39 . 6 PRO CB C 31 0.2 . 40 . 6 PRO HB2 H 1.9 0.02 . 41 . 6 PRO HB3 H 2.33 0.02 . 42 . 6 PRO HG2 H 2.01 0.02 . 43 . 6 PRO HG3 H 2.09 0.02 . 44 . 6 PRO HD2 H 3.73 0.02 . 45 . 6 PRO HD3 H 3.81 0.02 . 46 . 7 ALA H H 8.22 0.02 . 47 . 7 ALA N N 123.15 0.2 . 48 . 7 ALA CA C 53.61 0.04 . 49 . 7 ALA HA H 4.26 0.02 . 50 . 7 ALA C C 179.44 0.2 . 51 . 7 ALA CB C 18.33 0.06 . 52 . 7 ALA HB H 1.45 0.02 . 53 . 8 ARG H H 8.37 0.02 . 54 . 8 ARG N N 120.9 0.2 . 55 . 8 ARG CA C 57.6 0.02 . 56 . 8 ARG HA H 4.2 0.02 . 57 . 8 ARG C C 177.76 0.2 . 58 . 8 ARG CB C 30.23 0.01 . 59 . 8 ARG HB2 H 1.87 0.02 . 60 . 8 ARG HG2 H 1.64 0.02 . 61 . 8 ARG HG3 H 1.75 0.02 . 62 . 8 ARG HD2 H 3.22 0.02 . 63 . 8 ARG HE H 7.3 0.02 . 64 . 8 ARG HH11 H 7.01 0.01 . 65 . 8 ARG HH21 H 6.57 0.02 . 66 . 9 ALA H H 8.32 0.02 . 67 . 9 ALA N N 124.52 0.2 . 68 . 9 ALA CA C 53.76 0.05 . 69 . 9 ALA HA H 4.25 0.02 . 70 . 9 ALA C C 178.98 0.2 . 71 . 9 ALA CB C 18.18 0.2 . 72 . 9 ALA HB H 1.46 0.02 . 73 . 10 ALA H H 8.24 0.02 . 74 . 10 ALA N N 122.55 0.2 . 75 . 10 ALA CA C 53.81 0.2 . 76 . 10 ALA HA H 4.21 0.02 . 77 . 10 ALA C C 179.46 0.2 . 78 . 10 ALA CB C 18.18 0.2 . 79 . 10 ALA HB H 1.47 0.02 . 80 . 11 ALA H H 8.2 0.02 . 81 . 11 ALA N N 122.43 0.2 . 82 . 11 ALA CA C 53.77 0.2 . 83 . 11 ALA HA H 4.25 0.02 . 84 . 11 ALA C C 179.51 0.2 . 85 . 11 ALA CB C 18.1 0.2 . 86 . 11 ALA HB H 1.47 0.02 . 87 . 12 LYS H H 8.1 0.02 . 88 . 12 LYS N N 122.09 0.2 . 89 . 12 LYS CA C 58 0.02 . 90 . 12 LYS HA H 4.31 0.02 . 91 . 12 LYS C C 177.53 0.2 . 92 . 12 LYS CB C 32.19 0.01 . 93 . 12 LYS HB2 H 1.88 0.02 . 94 . 12 LYS HB3 H 2.01 0.02 . 95 . 12 LYS HG2 H 1.49 0.02 . 96 . 12 LYS HD2 H 1.65 0.02 . 97 . 12 LYS HD3 H 1.75 0.01 . 98 . 12 LYS HE2 H 3.02 0.02 . 99 . 12 LYS HZ H 7.8 0.02 . 100 . 13 THR H H 8.33 0.02 . 101 . 13 THR N N 116.11 0.2 . 102 . 13 THR CA C 64.91 0.2 . 103 . 13 THR HA H 4.13 0.02 . 104 . 13 THR C C 176.56 0.2 . 105 . 13 THR CB C 69.25 0.2 . 106 . 13 THR HB H 4.32 0.02 . 107 . 13 THR HG2 H 1.29 0.02 . 108 . 14 ALA H H 8.17 0.02 . 109 . 14 ALA N N 124.16 0.2 . 110 . 14 ALA CA C 54.69 0.2 . 111 . 14 ALA HA H 4.26 0.02 . 112 . 14 ALA C C 179.94 0.2 . 113 . 14 ALA CB C 17.86 0.2 . 114 . 14 ALA HB H 1.49 0.02 . 115 . 15 ALA H H 8.29 0.02 . 116 . 15 ALA N N 122.96 0.2 . 117 . 15 ALA CA C 54.93 0.2 . 118 . 15 ALA HA H 4.21 0.02 . 119 . 15 ALA C C 181.01 0.2 . 120 . 15 ALA CB C 17.82 0.03 . 121 . 15 ALA HB H 1.53 0.02 . 122 . 16 ASP H H 8.72 0.02 . 123 . 16 ASP N N 121.39 0.2 . 124 . 16 ASP CA C 56.54 0.2 . 125 . 16 ASP HA H 4.46 0.02 . 126 . 16 ASP C C 178.86 0.2 . 127 . 16 ASP CB C 39.15 0.2 . 128 . 16 ASP HB2 H 2.69 0.02 . 129 . 16 ASP HB3 H 2.87 0.02 . 130 . 17 ALA H H 8.04 0.02 . 131 . 17 ALA N N 124.3 0.2 . 132 . 17 ALA CA C 54.73 0.04 . 133 . 17 ALA HA H 4.24 0.02 . 134 . 17 ALA C C 180.6 0.2 . 135 . 17 ALA CB C 17.48 0.02 . 136 . 17 ALA HB H 1.52 0.02 . 137 . 18 LEU H H 7.97 0.02 . 138 . 18 LEU N N 121.62 0.2 . 139 . 18 LEU CA C 57.68 0.06 . 140 . 18 LEU HA H 4.27 0.02 . 141 . 18 LEU C C 179.05 0.2 . 142 . 18 LEU CB C 41.54 0.2 . 143 . 18 LEU HB2 H 1.79 0.02 . 144 . 18 LEU HD1 H 0.93 0.02 . 145 . 18 LEU HD2 H 0.97 0.02 . 146 . 18 LEU HG H 1.71 0.02 . 147 . 19 ALA H H 7.96 0.02 . 148 . 19 ALA N N 121.95 0.2 . 149 . 19 ALA CA C 54.64 0.2 . 150 . 19 ALA HA H 4.14 0.02 . 151 . 19 ALA C C 180.72 0.2 . 152 . 19 ALA CB C 17.66 0.2 . 153 . 19 ALA HB H 1.54 0.02 . 154 . 20 ALA H H 8.11 0.02 . 155 . 20 ALA N N 121.98 0.2 . 156 . 20 ALA CA C 54.58 0.2 . 157 . 20 ALA HA H 4.26 0.02 . 158 . 20 ALA C C 179.78 0.2 . 159 . 20 ALA CB C 17.54 0.2 . 160 . 20 ALA HB H 1.52 0.02 . 161 . 21 ALA H H 7.96 0.02 . 162 . 21 ALA N N 122.33 0.2 . 163 . 21 ALA CA C 54.58 0.2 . 164 . 21 ALA HA H 4.21 0.02 . 165 . 21 ALA C C 180.72 0.2 . 166 . 21 ALA CB C 17.45 0.04 . 167 . 21 ALA HB H 1.56 0.02 . 168 . 22 LYS H H 8.15 0.02 . 169 . 22 LYS N N 120.06 0.2 . 170 . 22 LYS CA C 59.26 0.2 . 171 . 22 LYS HA H 4.13 0.02 . 172 . 22 LYS C C 179 0.2 . 173 . 22 LYS CB C 32.47 0.2 . 174 . 22 LYS HB2 H 1.93 0.02 . 175 . 22 LYS HG2 H 1.44 0.01 . 176 . 22 LYS HD2 H 1.68 0.02 . 177 . 22 LYS HE2 H 2.96 0.02 . 178 . 22 LYS HZ H 7.71 0.02 . 179 . 23 LYS H H 7.92 0.02 . 180 . 23 LYS N N 122.45 0.2 . 181 . 23 LYS CA C 58.94 0.2 . 182 . 23 LYS HA H 4.25 0.02 . 183 . 23 LYS C C 177.77 0.2 . 184 . 23 LYS CB C 32.02 0.04 . 185 . 23 LYS HB2 H 1.9 0.02 . 186 . 23 LYS HB3 H 2.06 0.02 . 187 . 23 LYS HG2 H 1.51 0.02 . 188 . 23 LYS HG3 H 1.45 0.02 . 189 . 23 LYS HD2 H 1.66 0.02 . 190 . 23 LYS HD3 H 1.76 0.01 . 191 . 23 LYS HE2 H 3.03 0.02 . 192 . 23 LYS HZ H 7.84 0.02 . 193 . 24 THR H H 8.41 0.02 . 194 . 24 THR N N 116.66 0.2 . 195 . 24 THR CA C 65.94 0.05 . 196 . 24 THR HA H 4.04 0.02 . 197 . 24 THR C C 176.9 0.2 . 198 . 24 THR CB C 68.65 0.05 . 199 . 24 THR HB H 4.25 0.02 . 200 . 24 THR HG2 H 1.29 0.02 . 201 . 25 ALA H H 7.92 0.02 . 202 . 25 ALA N N 123.53 0.2 . 203 . 25 ALA CA C 54.53 0.2 . 204 . 25 ALA HA H 4.27 0.02 . 205 . 25 ALA C C 179.99 0.2 . 206 . 25 ALA CB C 17.7 0.2 . 207 . 25 ALA HB H 1.5 0.02 . 208 . 26 ALA H H 8.27 0.02 . 209 . 26 ALA N N 123.96 0.2 . 210 . 26 ALA CA C 54.66 0.07 . 211 . 26 ALA HA H 4.25 0.02 . 212 . 26 ALA C C 180.79 0.2 . 213 . 26 ALA CB C 17.74 0.2 . 214 . 26 ALA HB H 1.55 0.02 . 215 . 27 ASP H H 8.82 0.02 . 216 . 27 ASP N N 121.26 0.2 . 217 . 27 ASP CA C 56.57 0.2 . 218 . 27 ASP HA H 4.45 0.02 . 219 . 27 ASP C C 178.81 0.2 . 220 . 27 ASP CB C 39.26 0.2 . 221 . 27 ASP HB2 H 2.68 0.02 . 222 . 27 ASP HB3 H 2.84 0.02 . 223 . 28 ALA H H 8 0.02 . 224 . 28 ALA N N 123.84 0.2 . 225 . 28 ALA CA C 54.45 0.2 . 226 . 28 ALA HA H 4.27 0.02 . 227 . 28 ALA C C 180.16 0.2 . 228 . 28 ALA CB C 17.46 0.2 . 229 . 28 ALA HB H 1.52 0.02 . 230 . 29 ALA H H 8.06 0.02 . 231 . 29 ALA N N 122.34 0.2 . 232 . 29 ALA CA C 54.13 0.2 . 233 . 29 ALA HA H 4.26 0.01 . 234 . 29 ALA C C 180.04 0.2 . 235 . 29 ALA CB C 17.52 0.2 . 236 . 29 ALA HB H 1.53 0.02 . 237 . 30 ALA H H 8.02 0.02 . 238 . 30 ALA N N 121.81 0.2 . 239 . 30 ALA CA C 54.21 0.2 . 240 . 30 ALA HA H 4.23 0.02 . 241 . 30 ALA C C 179.73 0.2 . 242 . 30 ALA CB C 17.46 0.2 . 243 . 30 ALA HB H 1.52 0.02 . 244 . 31 ALA H H 7.9 0.02 . 245 . 31 ALA N N 121.28 0.2 . 246 . 31 ALA CA C 53.73 0.2 . 247 . 31 ALA HA H 4.22 0.02 . 248 . 31 ALA C C 179.31 0.2 . 249 . 31 ALA CB C 17.94 0.2 . 250 . 31 ALA HB H 1.48 0.02 . 251 . 32 ALA H H 7.82 0.02 . 252 . 32 ALA N N 120.78 0.2 . 253 . 32 ALA CA C 53.57 0.2 . 254 . 32 ALA HA H 4.24 0.02 . 255 . 32 ALA C C 178.56 0.2 . 256 . 32 ALA CB C 18.02 0.2 . 257 . 32 ALA HB H 1.47 0.02 . 258 . 33 ALA H H 7.72 0.02 . 259 . 33 ALA N N 120.73 0.2 . 260 . 33 ALA CA C 52.69 0.2 . 261 . 33 ALA HA H 4.28 0.02 . 262 . 33 ALA C C 177.48 0.2 . 263 . 33 ALA CB C 18.43 0.2 . 264 . 33 ALA HB H 1.46 0.02 . 265 . 34 ALA H H 7.67 0.02 . 266 . 34 ALA N N 122.07 0.2 . 267 . 34 ALA CA C 52.03 0.01 . 268 . 34 ALA HA H 4.34 0.02 . 269 . 34 ALA C C 176.38 0.2 . 270 . 34 ALA CB C 18.83 0.2 . 271 . 34 ALA HB H 1.44 0.02 . 272 . 35 ALA H H 7.65 0.02 . 273 . 35 ALA N N 128.66 0.2 . 274 . 35 ALA CA C 53.82 0.2 . 275 . 35 ALA HA H 4.08 0.02 . 276 . 35 ALA CB C 19.34 0.2 . 277 . 35 ALA HB H 1.38 0.02 . stop_ save_ save_chemical_shift_set_278K _Saveframe_category assigned_chemical_shifts _Details . loop_ _Sample_label $sample_1 $sample_2 stop_ _Sample_conditions_label $sample_cond_2 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name rSS3 _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 GLY CA C 42.91 0.2 . 2 . 1 GLY C C 169.92 0.2 . 3 . 2 SER H H 8.8 0.02 . 4 . 2 SER N N 114.14 0.2 . 5 . 2 SER CA C 58.05 0.2 . 6 . 2 SER HA H 4.5 0.02 . 7 . 2 SER C C 174.4 0.2 . 8 . 2 SER CB C 63.69 0.03 . 9 . 2 SER HB2 H 3.87 0.01 . 10 . 3 MET H H 8.74 0.02 . 11 . 3 MET N N 121.06 0.2 . 12 . 3 MET CA C 55.4 0.02 . 13 . 3 MET HA H 4.52 0.02 . 14 . 3 MET C C 175.72 0.2 . 15 . 3 MET CB C 32.39 0.02 . 16 . 3 MET HB2 H 2.09 0.02 . 17 . 3 MET HB3 H 2.01 0.02 . 18 . 3 MET HG2 H 2.61 0.02 . 19 . 3 MET HG3 H 2.55 0.01 . 20 . 4 ASN H H 8.54 0.02 . 21 . 4 ASN N N 118.83 0.2 . 22 . 4 ASN CA C 52.64 0.02 . 23 . 4 ASN HA H 4.7 0.02 . 24 . 4 ASN C C 174.25 0.2 . 25 . 4 ASN CB C 38.37 0.01 . 26 . 4 ASN HB2 H 2.83 0.02 . 27 . 4 ASN HB3 H 2.78 0.02 . 28 . 4 ASN HD21 H 7.73 0.02 . 29 . 4 ASN HD22 H 7.02 0.01 . 30 . 5 ALA H H 8.47 0.02 . 31 . 5 ALA N N 123.52 0.2 . 32 . 5 ALA CA C 51.89 0.2 . 33 . 5 ALA HA H 4.51 0.02 . 34 . 5 ALA CB C 17.13 0.2 . 35 . 5 ALA HB H 1.39 0.01 . 36 . 6 PRO CA C 63.4 0.2 . 37 . 6 PRO HA H 4.41 0.02 . 38 . 6 PRO C C 177.28 0.2 . 39 . 6 PRO CB C 31.31 0.2 . 40 . 6 PRO HB2 H 2.32 0.02 . 41 . 6 PRO HB3 H 1.89 0.01 . 42 . 6 PRO HG2 H 2.06 0.01 . 43 . 6 PRO HG3 H 2.01 0.01 . 44 . 6 PRO HD2 H 3.8 0.02 . 45 . 6 PRO HD3 H 3.68 0.01 . 46 . 7 ALA H H 8.41 0.02 . 47 . 7 ALA N N 122.26 0.01 . 48 . 7 ALA CA C 52.72 0.2 . 49 . 7 ALA HA H 4.24 0.02 . 50 . 7 ALA C C 178.54 0.2 . 51 . 7 ALA CB C 18.37 0.01 . 52 . 7 ALA HB H 1.42 0.02 . 53 . 8 ARG H H 8.44 0.02 . 54 . 8 ARG N N 119.3 0.2 . 55 . 8 ARG CA C 56.76 0.2 . 56 . 8 ARG HA H 4.24 0.02 . 57 . 8 ARG C C 176.91 0.2 . 58 . 8 ARG CB C 30.25 0.03 . 59 . 8 ARG HB2 H 1.83 0.01 . 60 . 8 ARG HG2 H 1.7 0.01 . 61 . 8 ARG HG3 H 1.63 0.01 . 62 . 8 ARG HD2 H 3.22 0.02 . 63 . 8 ARG HE H 7.26 0.02 . 64 . 8 ARG HH11 H 6.97 0.02 . 65 . 8 ARG HH21 H 6.53 0.02 . 66 . 9 ALA H H 8.43 0.02 . 67 . 9 ALA N N 123.52 0.2 . 68 . 9 ALA CA C 53.03 0.2 . 69 . 9 ALA HA H 4.26 0.02 . 70 . 9 ALA C C 178.24 0.2 . 71 . 9 ALA CB C 18.09 0.05 . 72 . 9 ALA HB H 1.42 0.02 . 73 . 10 ALA H H 8.37 0.02 . 74 . 10 ALA N N 121.64 0.2 . 75 . 10 ALA CA C 54.37 0.03 . 76 . 10 ALA HA H 4.23 0.02 . 77 . 10 ALA C C 178.58 0.2 . 78 . 10 ALA CB C 18.2 0.02 . 79 . 10 ALA HB H 1.44 0.02 . 80 . 11 ALA H H 8.34 0.02 . 81 . 11 ALA N N 121.53 0.01 . 82 . 11 ALA CA C 53.18 0.05 . 83 . 11 ALA HA H 4.25 0.02 . 84 . 11 ALA C C 178.88 0.2 . 85 . 11 ALA CB C 18.17 0.08 . 86 . 11 ALA HB H 1.44 0.02 . 87 . 12 LYS H H 8.31 0.02 . 88 . 12 LYS N N 120.16 0.2 . 89 . 12 LYS CA C 57.41 0.02 . 90 . 12 LYS HA H 4.34 0.02 . 91 . 12 LYS C C 177.21 0.2 . 92 . 12 LYS CB C 32.35 0.2 . 93 . 12 LYS HB2 H 1.96 0.02 . 94 . 12 LYS HB3 H 1.85 0.01 . 95 . 12 LYS HG2 H 1.51 0.02 . 96 . 12 LYS HG3 H 1.47 0.01 . 97 . 12 LYS HD2 H 1.72 0.02 . 98 . 12 LYS HD3 H 1.66 0.01 . 99 . 12 LYS HE2 H 3.01 0.02 . 100 . 12 LYS HZ H 7.72 0.02 . 101 . 13 THR H H 8.41 0.02 . 102 . 13 THR N N 113.96 0.2 . 103 . 13 THR CA C 63.43 0.06 . 104 . 13 THR HA H 4.22 0.01 . 105 . 13 THR C C 175.88 0.2 . 106 . 13 THR CB C 69.68 0.02 . 107 . 13 THR HB H 4.34 0.02 . 108 . 13 THR HG2 H 1.28 0.01 . 109 . 14 ALA H H 8.41 0.02 . 110 . 14 ALA N N 123.36 0.2 . 111 . 14 ALA CA C 54.14 0.01 . 112 . 14 ALA C C 179.45 0.2 . 113 . 14 ALA CB C 18.22 0.01 . 114 . 15 ALA H H 8.38 0.02 . 115 . 15 ALA N N 121.17 0.2 . 116 . 15 ALA CA C 54.45 0.01 . 117 . 15 ALA HA H 4.21 0.02 . 118 . 15 ALA C C 180.22 0.2 . 119 . 15 ALA CB C 17.84 0.04 . 120 . 16 ASP H H 8.57 0.02 . 121 . 16 ASP N N 118.83 0.2 . 122 . 16 ASP CA C 56.17 0.2 . 123 . 16 ASP HA H 4.48 0.02 . 124 . 16 ASP C C 178.27 0.2 . 125 . 16 ASP CB C 39.52 0.01 . 126 . 16 ASP HB2 H 2.84 0.01 . 127 . 16 ASP HB3 H 2.7 0.01 . 128 . 17 ALA H H 8.17 0.02 . 129 . 17 ALA N N 123.05 0.2 . 130 . 17 ALA CA C 54.32 0.02 . 131 . 17 ALA HA H 4.25 0.02 . 132 . 17 ALA C C 180.13 0.2 . 133 . 17 ALA CB C 17.79 0.01 . 134 . 17 ALA HB H 1.52 0.02 . 135 . 18 LEU H H 8.08 0.02 . 136 . 18 LEU N N 119.77 0.2 . 137 . 18 LEU CA C 57.1 0.04 . 138 . 18 LEU HA H 4.25 0.02 . 139 . 18 LEU C C 178.83 0.2 . 140 . 18 LEU CB C 41.41 0.2 . 141 . 18 LEU HB2 H 1.78 0.01 . 142 . 18 LEU HB3 H 1.74 0.01 . 143 . 18 LEU HD1 H 0.95 0.02 . 144 . 18 LEU HD2 H 0.91 0.02 . 145 . 18 LEU HG H 1.7 0.01 . 146 . 19 ALA H H 8.04 0.01 . 147 . 19 ALA N N 120.76 0.2 . 148 . 19 ALA CA C 54.24 0.01 . 149 . 19 ALA HA H 4.14 0.02 . 150 . 19 ALA C C 180.13 0.2 . 151 . 19 ALA CB C 17.84 0.03 . 152 . 19 ALA HB H 1.52 0.02 . 153 . 20 ALA H H 8.16 0.02 . 154 . 20 ALA N N 120.23 0.2 . 155 . 20 ALA CA C 53.97 0.02 . 156 . 20 ALA HA H 4.23 0.02 . 157 . 20 ALA C C 179.32 0.2 . 158 . 20 ALA CB C 17.92 0.03 . 159 . 20 ALA HB H 1.5 0.02 . 160 . 21 ALA H H 8.02 0.02 . 161 . 21 ALA N N 120.51 0.2 . 162 . 21 ALA CA C 54.05 0.2 . 163 . 21 ALA HA H 4.23 0.02 . 164 . 21 ALA C C 180.01 0.2 . 165 . 21 ALA CB C 17.79 0.2 . 166 . 21 ALA HB H 1.54 0.02 . 167 . 22 LYS H H 8.2 0.02 . 168 . 22 LYS N N 118.36 0.2 . 169 . 22 LYS CA C 58.25 0.05 . 170 . 22 LYS HA H 4.15 0.02 . 171 . 22 LYS C C 178.41 0.2 . 172 . 22 LYS CB C 32.19 0.05 . 173 . 22 LYS HB2 H 1.91 0.01 . 174 . 22 LYS HG2 H 1.62 0.01 . 175 . 22 LYS HG3 H 1.44 0.01 . 176 . 22 LYS HD2 H 1.68 0.02 . 177 . 22 LYS HE2 H 2.97 0.02 . 178 . 22 LYS HZ H 7.66 0.02 . 179 . 23 LYS H H 8.07 0.02 . 180 . 23 LYS N N 120.93 0.2 . 181 . 23 LYS CA C 58.02 0.05 . 182 . 23 LYS HA H 4.26 0.02 . 183 . 23 LYS C C 177.5 0.2 . 184 . 23 LYS CB C 32.13 0.02 . 185 . 23 LYS HB2 H 2.02 0.02 . 186 . 23 LYS HB3 H 1.89 0.01 . 187 . 23 LYS HG2 H 1.48 0.02 . 188 . 23 LYS HD2 H 1.74 0.01 . 189 . 23 LYS HD3 H 1.66 0.01 . 190 . 23 LYS HE2 H 3.02 0.02 . 191 . 23 LYS HZ H 7.76 0.02 . 192 . 24 THR H H 8.45 0.02 . 193 . 24 THR N N 114.61 0.2 . 194 . 24 THR CA C 65.05 0.02 . 195 . 24 THR HA H 4.1 0.01 . 196 . 24 THR C C 176.24 0.2 . 197 . 24 THR CB C 69.09 0.2 . 198 . 24 THR HB H 4.26 0.02 . 199 . 24 THR HG2 H 1.28 0.02 . 200 . 25 ALA H H 8.11 0.02 . 201 . 25 ALA N N 122.51 0.2 . 202 . 25 ALA CA C 54.19 0.2 . 203 . 25 ALA HA H 4.27 0.02 . 204 . 25 ALA C C 179.5 0.2 . 205 . 25 ALA CB C 17.87 0.03 . 206 . 25 ALA HB H 1.49 0.02 . 207 . 26 ALA H H 8.33 0.02 . 208 . 26 ALA N N 122.04 0.02 . 209 . 26 ALA CA C 54.27 0.08 . 210 . 26 ALA HA H 4.25 0.02 . 211 . 26 ALA C C 180.01 0.2 . 212 . 26 ALA CB C 17.88 0.2 . 213 . 26 ALA HB H 1.51 0.02 . 214 . 27 ASP H H 8.7 0.02 . 215 . 27 ASP N N 119.14 0.2 . 216 . 27 ASP CA C 55.99 0.2 . 217 . 27 ASP HA H 4.46 0.02 . 218 . 27 ASP C C 178.17 0.2 . 219 . 27 ASP CB C 39.47 0.02 . 220 . 27 ASP HB2 H 2.81 0.01 . 221 . 27 ASP HB3 H 2.69 0.01 . 222 . 28 ALA H H 8.12 0.02 . 223 . 28 ALA N N 122.58 0.2 . 224 . 28 ALA CA C 54.15 0.01 . 225 . 28 ALA HA H 4.25 0.02 . 226 . 28 ALA C C 179.53 0.2 . 227 . 28 ALA CB C 17.87 0.02 . 228 . 28 ALA HB H 1.5 0.02 . 229 . 29 ALA H H 8.13 0.02 . 230 . 29 ALA N N 120.76 0.2 . 231 . 29 ALA CA C 53.92 0.03 . 232 . 29 ALA HA H 4.24 0.02 . 233 . 29 ALA C C 179.43 0.2 . 234 . 29 ALA CB C 17.76 0.04 . 235 . 29 ALA HB H 1.49 0.02 . 236 . 30 ALA H H 8.08 0.02 . 237 . 30 ALA N N 120.47 0.2 . 238 . 30 ALA CA C 53.7 0.05 . 239 . 30 ALA HA H 4.25 0.02 . 240 . 30 ALA C C 179.15 0.2 . 241 . 30 ALA CB C 18.02 0.02 . 242 . 30 ALA HB H 1.49 0.02 . 243 . 31 ALA H H 8.01 0.02 . 244 . 31 ALA N N 119.99 0.2 . 245 . 31 ALA CA C 53.18 0.04 . 246 . 31 ALA HA H 4.22 0.02 . 247 . 31 ALA C C 178.73 0.2 . 248 . 31 ALA CB C 17.96 0.03 . 249 . 31 ALA HB H 1.45 0.02 . 250 . 32 ALA H H 7.96 0.02 . 251 . 32 ALA N N 119.77 0.2 . 252 . 32 ALA CA C 52.88 0.04 . 253 . 32 ALA HA H 4.25 0.02 . 254 . 32 ALA C C 178.06 0.2 . 255 . 32 ALA CB C 18.34 0.2 . 256 . 32 ALA HB H 1.46 0.02 . 257 . 33 ALA H H 7.91 0.02 . 258 . 33 ALA N N 120.08 0.2 . 259 . 33 ALA CA C 52.37 0.02 . 260 . 33 ALA C C 177.29 0.2 . 261 . 33 ALA CB C 18.61 0.05 . 262 . 33 ALA HB H 1.45 0.02 . 263 . 34 ALA H H 7.92 0.02 . 264 . 34 ALA N N 121.26 0.2 . 265 . 34 ALA CA C 52.05 0.02 . 266 . 34 ALA HA H 4.32 0.02 . 267 . 34 ALA C C 176.26 0.2 . 268 . 34 ALA CB C 18.82 0.2 . 269 . 34 ALA HB H 1.43 0.02 . 270 . 35 ALA H H 7.83 0.02 . 271 . 35 ALA N N 127.38 0.2 . 272 . 35 ALA CA C 53.74 0.2 . 273 . 35 ALA HA H 4.1 0.02 . 274 . 35 ALA CB C 18.86 0.2 . 275 . 35 ALA HB H 1.36 0.02 . stop_ save_ save_rdc_nh _Saveframe_category residual_dipolar_couplings loop_ _Sample_label $sample_3 stop_ _Details . _Sample_conditions_label $sample_cond_2 _Spectrometer_frequency_1H 600 _Text_data_format . _Text_data . loop_ _Residual_dipolar_coupling_ID _Atom_one_residue_seq_code _Atom_one_residue_label _Atom_one_atom_name _Atom_two_residue_seq_code _Atom_two_residue_label _Atom_two_atom_name _Residual_dipolar_coupling_value _Atom_one_mol_system_component_name _Atom_two_mol_system_component_name _Residual_dipolar_coupling_min_value _Residual_dipolar_coupling_max_value _Residual_dipolar_coupling_value_error 1DHN 13 THR H 13 THR N -6.600 ? ? . . . 1DHN 14 ALA H 14 ALA N -7.600 ? ? . . . 1DHN 15 ALA H 15 ALA N -8.600 ? ? . . . 1DHN 16 ASP H 16 ASP N -6.600 ? ? . . . 1DHN 17 ALA H 17 ALA N -7.700 ? ? . . . 1DHN 18 LEU H 18 LEU N -8.700 ? ? . . . 1DHN 19 ALA H 19 ALA N -9.700 ? ? . . . 1DHN 20 ALA H 20 ALA N -7.600 ? ? . . . 1DHN 21 ALA H 21 ALA N -9.500 ? ? . . . 1DHN 22 LYS H 22 LYS N -6.600 ? ? . . . 1DHN 23 LYS H 23 LYS N -8.500 ? ? . . . 1DHN 24 THR H 24 THR N -8.600 ? ? . . . 1DHN 25 ALA H 25 ALA N -11.40 ? ? . . . 1DHN 26 ALA H 26 ALA N -9.500 ? ? . . . 1DHN 27 ASP H 27 ASP N -8.600 ? ? . . . 1DHN 28 ALA H 28 ALA N -7.600 ? ? . . . 1DHN 29 ALA H 29 ALA N -8.500 ? ? . . . 1DHN 30 ALA H 30 ALA N -7.600 ? ? . . . stop_ save_