data_5120 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H Chemical Shift Assigments for the mEGF/TGFalpha44-50 chimeric growth factor ; _BMRB_accession_number 5120 _BMRB_flat_file_name bmr5120.str _Entry_type original _Submission_date 2001-08-23 _Accession_date 2001-08-23 _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 Chamberlin S. G. . 2 Brennan L. . . 3 Puddicombe S. M. . 4 Davies D. E. . 5 Turner D. L. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 232 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2002-01-25 original BMRB . stop_ _Original_release_date 2001-08-23 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Solution Structure of the mEGF/TGFalpha44-50 Chimeric Growth Factor ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code 21590355 _PubMed_ID 11733021 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Chamberlin S. G. . 2 Brennan L. . . 3 Puddicombe S. M. . 4 Davies D. E. . 5 Turner D. L. . stop_ _Journal_abbreviation 'Eur. J. Biochem.' _Journal_volume 268 _Journal_issue 23 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 6247 _Page_last 6255 _Year 2001 _Details . loop_ _Keyword 'EGF/TGFalpha44-50 chimera' NMR stop_ save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Puddicombe SM, Wood L, Chamberlin SG, Davies DE. The interaction of an epidermal growth factor/transforming growth facotr alpha tail chimera with the human epidermal growth factor receptor reveals uncexpected complexities. The Journal of Biological Chemistry. 271, 30392-30397. ; _Citation_title ; The interaction of an epidermal growth factor/transforming growth factor alpha tail chimera with the human epidermal growth factor receptor reveals unexpected complexities. ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 8940002 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Puddicombe 'S M' M. . 2 Wood L . . 3 Chamberlin 'S G' G. . 4 Davies 'D E' E. . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'The Journal of biological chemistry' _Journal_volume 271 _Journal_issue 48 _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 30392 _Page_last 30397 _Year 1996 _Details ; It has been assumed that substitution of homologous regions of transforming growth factor alpha (TGF-alpha) into epidermal growth factor (EGF) can be used to probe ligand-receptor recognition without detrimental effects on ligand characteristics for the human EGF receptor (EGFR). We show that a chimera of murine (m) EGF in which the carboxyl-terminal tail is substituted for that of TGF-alpha (mEGF/TGF-alpha44-50) results in complex features that belie this initial simplistic assumption. Comparison of EGF and mEGF/TGF-alpha44-50 in equilibrium binding assays showed that although the relative binding affinity of the chimera was reduced 80-200-fold, it was more potent than EGF in mitogenesis assays using NR6/HER cells. This superagonist activity could not be attributed to differences in ligand processing or to binding to other members of the c-erbB family. It appeared to be due, in part, to choice of an EGFR-overexpressing target cell where high receptor number compensated for the low affinity of the ligand; it also appeared to be related to the ability of the chimera to activate the EGFR tyrosine kinase. Thus, when EGFR autophosphorylation was measured, mEGF/TGF-alpha44-50 was more potent than EGF, despite its low affinity. When tested using chicken embryo fibroblasts, substitution of the TGF-alpha carboxyl-terminal tail into mEGF failed to enhance its binding affinity for chicken EGFRs; however, the chimera was intermediate in potency between TGF-alpha and mEGF in mitogenesis assays. Our results suggest a contextual requirement for EGFR recognition which is ligand-specific. Further, the unpredictable responses to chimeric ligands underline the complex nature of the processes of ligand recognition, receptor activation, and the ensuing cellular response. ; save_ ################################## # Molecular system description # ################################## save_system_mEGF_TGFalpha44-50_chimera _Saveframe_category molecular_system _Mol_system_name 'mEGF/TGFalpha44-50 chimera' _Abbreviation_common 'mEGF/TGFalpha44-50 chimera' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'mEGF/TGFalpha44-50 chimera' $chimera stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all disulfide bound' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_chimera _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'mEGF/TGFalpha44-50 chimera' _Abbreviation_common 'mEGF/TGFalpha44-50 chimera' _Molecular_mass 5000 _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 49 _Mol_residue_sequence ; NSYPGCPSSYDGYCLNGGVC MHIESLDSYTCNCVIGYSGD RCEHADLLA ; loop_ _Residue_seq_code _Residue_label 1 ASN 2 SER 3 TYR 4 PRO 5 GLY 6 CYS 7 PRO 8 SER 9 SER 10 TYR 11 ASP 12 GLY 13 TYR 14 CYS 15 LEU 16 ASN 17 GLY 18 GLY 19 VAL 20 CYS 21 MET 22 HIS 23 ILE 24 GLU 25 SER 26 LEU 27 ASP 28 SER 29 TYR 30 THR 31 CYS 32 ASN 33 CYS 34 VAL 35 ILE 36 GLY 37 TYR 38 SER 39 GLY 40 ASP 41 ARG 42 CYS 43 GLU 44 HIS 45 ALA 46 ASP 47 LEU 48 LEU 49 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 1GK5 'Solution Structure The MegfTGFALPHA44-50 Chimeric Growth Factor' 97.96 49 100.00 100.00 2.18e-19 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $chimera 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 $chimera '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 _Isotopic_labeling $chimera 3 mM . stop_ save_ ############################ # Computer software used # ############################ save_XEASY _Saveframe_category software _Name XEASY _Version . _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer _Saveframe_category NMR_spectrometer _Manufacturer VARIAN _Model VXR _Field_strength 500 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D-1H-NOESY_1 _Saveframe_category NMR_applied_experiment _Experiment_name 2D-1H-NOESY _Sample_label . save_ save_2D-1H-TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name 2D-1H-TOCSY _Sample_label . save_ save_2D-1H-COSY_3 _Saveframe_category NMR_applied_experiment _Experiment_name 2D-1H-COSY _Sample_label . save_ ####################### # Sample conditions # ####################### save_sample_cond_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 3.0 0.1 n/a temperature 298 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 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_ _Experiment_label 2D-1H-NOESY 2D-1H-TOCSY 2D-1H-COSY stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'mEGF/TGFalpha44-50 chimera' _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 ASN HA H 4.32 0.01 1 2 . 1 ASN HB3 H 2.83 0.01 2 3 . 1 ASN HD21 H 7.58 0.01 2 4 . 1 ASN HD22 H 7.00 0.01 2 5 . 2 SER H H 8.58 0.01 1 6 . 2 SER HA H 4.60 0.01 1 7 . 2 SER HB2 H 3.72 0.01 2 8 . 2 SER HB3 H 3.68 0.01 2 9 . 3 TYR H H 8.36 0.01 1 10 . 3 TYR HA H 4.90 0.01 1 11 . 3 TYR HB2 H 3.06 0.01 2 12 . 3 TYR HB3 H 2.71 0.01 2 13 . 3 TYR HD1 H 7.12 0.01 1 14 . 3 TYR HE1 H 6.78 0.01 1 15 . 4 PRO HA H 4.65 0.01 1 16 . 4 PRO HB2 H 2.26 0.01 2 17 . 4 PRO HB3 H 2.10 0.01 2 18 . 4 PRO HG2 H 2.07 0.01 2 19 . 4 PRO HG3 H 2.00 0.01 2 20 . 4 PRO HD2 H 3.82 0.01 2 21 . 4 PRO HD3 H 3.63 0.01 2 22 . 5 GLY H H 8.10 0.01 1 23 . 5 GLY HA2 H 4.21 0.01 2 24 . 5 GLY HA3 H 4.07 0.01 2 25 . 6 CYS H H 8.70 0.01 1 26 . 6 CYS HA H 4.50 0.01 1 27 . 6 CYS HB2 H 2.95 0.01 2 28 . 6 CYS HB3 H 2.79 0.01 2 29 . 7 PRO HA H 4.57 0.01 1 30 . 7 PRO HB2 H 2.41 0.01 2 31 . 7 PRO HB3 H 2.08 0.01 2 32 . 7 PRO HG2 H 1.92 0.01 2 33 . 7 PRO HG3 H 1.84 0.01 2 34 . 7 PRO HD2 H 3.37 0.01 2 35 . 7 PRO HD3 H 2.69 0.01 2 36 . 8 SER H H 9.04 0.01 1 37 . 8 SER HA H 4.08 0.01 1 38 . 8 SER HB3 H 3.93 0.01 2 39 . 9 SER H H 8.02 0.01 1 40 . 9 SER HA H 4.13 0.01 1 41 . 9 SER HB3 H 3.76 0.01 2 42 . 10 TYR H H 8.09 0.01 1 43 . 10 TYR HA H 4.14 0.01 1 44 . 10 TYR HB2 H 3.26 0.01 2 45 . 10 TYR HB3 H 2.66 0.01 2 46 . 10 TYR HD1 H 6.41 0.01 1 47 . 10 TYR HE1 H 5.94 0.01 1 48 . 11 ASP H H 7.91 0.01 1 49 . 11 ASP HA H 4.47 0.01 1 50 . 11 ASP HB3 H 2.95 0.01 2 51 . 12 GLY H H 8.81 0.01 1 52 . 12 GLY HA2 H 3.96 0.01 2 53 . 12 GLY HA3 H 3.88 0.01 2 54 . 13 TYR H H 7.96 0.01 1 55 . 13 TYR HA H 4.13 0.01 1 56 . 13 TYR HB2 H 3.11 0.01 2 57 . 13 TYR HB3 H 3.02 0.01 2 58 . 13 TYR HD1 H 7.06 0.01 1 59 . 13 TYR HE1 H 6.77 0.01 1 60 . 13 TYR HE2 H 6.78 0.01 1 61 . 13 TYR HD2 H 7.06 0.01 1 62 . 14 CYS H H 8.79 0.01 1 63 . 14 CYS HA H 4.36 0.01 1 64 . 14 CYS HB2 H 2.60 0.01 2 65 . 14 CYS HB3 H 2.23 0.01 2 66 . 15 LEU H H 8.14 0.01 1 67 . 15 LEU HA H 4.26 0.01 1 68 . 15 LEU HB2 H 1.56 0.01 2 69 . 15 LEU HB3 H 1.27 0.01 2 70 . 15 LEU HG H 1.60 0.01 1 71 . 15 LEU HD1 H 0.71 0.01 2 72 . 16 ASN H H 9.01 0.01 1 73 . 16 ASN HA H 3.99 0.01 1 74 . 16 ASN HB2 H 1.93 0.01 2 75 . 16 ASN HB3 H 1.29 0.01 2 76 . 16 ASN HD21 H 7.81 0.01 2 77 . 16 ASN HD22 H 7.21 0.01 2 78 . 17 GLY H H 8.73 0.01 1 79 . 17 GLY HA2 H 4.00 0.01 2 80 . 17 GLY HA3 H 3.59 0.01 2 81 . 18 GLY H H 7.42 0.01 1 82 . 18 GLY HA2 H 4.31 0.01 2 83 . 18 GLY HA3 H 3.40 0.01 2 84 . 19 VAL H H 8.10 0.01 1 85 . 19 VAL HA H 4.32 0.01 1 86 . 19 VAL HB H 1.99 0.01 1 87 . 19 VAL HG2 H 0.99 0.01 2 88 . 20 CYS H H 8.88 0.01 1 89 . 20 CYS HA H 4.99 0.01 1 90 . 20 CYS HB2 H 3.36 0.01 2 91 . 20 CYS HB3 H 3.11 0.01 2 92 . 21 MET H H 9.60 0.01 1 93 . 21 MET HA H 4.97 0.01 1 94 . 21 MET HB3 H 1.89 0.01 2 95 . 21 MET HG3 H 2.43 0.01 2 96 . 21 MET HE H 1.92 0.01 1 97 . 22 HIS H H 8.80 0.01 1 98 . 22 HIS HA H 5.15 0.01 1 99 . 22 HIS HB2 H 3.17 0.01 2 100 . 22 HIS HB3 H 2.90 0.01 2 101 . 22 HIS HD2 H 6.53 0.01 1 102 . 22 HIS HE1 H 8.51 0.01 1 103 . 23 ILE H H 8.36 0.01 1 104 . 23 ILE HA H 4.06 0.01 1 105 . 23 ILE HB H 1.79 0.01 1 106 . 23 ILE HG2 H 0.80 0.01 1 107 . 23 ILE HG12 H 1.25 0.01 2 108 . 23 ILE HG13 H 0.91 0.01 2 109 . 23 ILE HD1 H 0.65 0.01 1 110 . 24 GLU H H 8.50 0.01 1 111 . 24 GLU HA H 3.65 0.01 1 112 . 24 GLU HB3 H 1.97 0.01 2 113 . 24 GLU HG3 H 2.34 0.01 2 114 . 25 SER H H 8.54 0.01 1 115 . 25 SER HA H 4.10 0.01 1 116 . 25 SER HB3 H 3.87 0.01 2 117 . 26 LEU H H 6.91 0.01 1 118 . 26 LEU HA H 4.40 0.01 1 119 . 26 LEU HB2 H 1.52 0.01 2 120 . 26 LEU HB3 H 1.42 0.01 2 121 . 26 LEU HG H 1.45 0.01 1 122 . 26 LEU HD1 H 0.82 0.01 2 123 . 26 LEU HD2 H 0.78 0.01 2 124 . 27 ASP H H 8.02 0.01 1 125 . 27 ASP HA H 4.34 0.01 1 126 . 27 ASP HB2 H 3.17 0.01 2 127 . 27 ASP HB3 H 2.71 0.01 2 128 . 28 SER H H 7.18 0.01 1 129 . 28 SER HA H 4.59 0.01 1 130 . 28 SER HB2 H 3.63 0.01 2 131 . 28 SER HB3 H 3.49 0.01 2 132 . 29 TYR H H 8.43 0.01 1 133 . 29 TYR HA H 5.23 0.01 1 134 . 29 TYR HB2 H 2.45 0.01 2 135 . 29 TYR HB3 H 2.26 0.01 2 136 . 29 TYR HD1 H 6.80 0.01 1 137 . 29 TYR HE1 H 6.23 0.01 1 138 . 29 TYR HE2 H 6.23 0.01 1 139 . 30 THR H H 9.00 0.01 1 140 . 30 THR HA H 4.98 0.01 1 141 . 30 THR HB H 4.11 0.01 1 142 . 30 THR HG2 H 1.07 0.01 1 143 . 31 CYS H H 8.63 0.01 1 144 . 31 CYS HA H 5.31 0.01 1 145 . 31 CYS HB2 H 2.77 0.01 2 146 . 31 CYS HB3 H 2.55 0.01 2 147 . 32 ASN H H 9.42 0.01 1 148 . 32 ASN HA H 5.00 0.01 1 149 . 32 ASN HB2 H 2.97 0.01 2 150 . 32 ASN HB3 H 2.77 0.01 2 151 . 32 ASN HD21 H 7.28 0.01 2 152 . 32 ASN HD22 H 6.81 0.01 2 153 . 33 CYS H H 8.94 0.01 1 154 . 33 CYS HA H 4.72 0.01 1 155 . 33 CYS HB2 H 3.28 0.01 2 156 . 33 CYS HB3 H 2.65 0.01 2 157 . 34 VAL H H 8.40 0.01 1 158 . 34 VAL HA H 4.09 0.01 1 159 . 34 VAL HB H 2.09 0.01 1 160 . 34 VAL HG1 H 1.15 0.01 2 161 . 34 VAL HG2 H 0.99 0.01 2 162 . 35 ILE H H 8.03 0.01 1 163 . 35 ILE HA H 3.93 0.01 1 164 . 35 ILE HB H 1.81 0.01 1 165 . 35 ILE HG2 H 0.76 0.01 1 166 . 35 ILE HG12 H 1.61 0.01 2 167 . 35 ILE HG13 H 1.28 0.01 2 168 . 35 ILE HD1 H 0.93 0.01 1 169 . 36 GLY H H 8.48 0.01 1 170 . 36 GLY HA2 H 4.22 0.01 2 171 . 36 GLY HA3 H 3.39 0.01 2 172 . 37 TYR H H 8.20 0.01 1 173 . 37 TYR HA H 5.35 0.01 1 174 . 37 TYR HB3 H 2.93 0.01 2 175 . 37 TYR HD1 H 6.83 0.01 1 176 . 37 TYR HE1 H 6.64 0.01 1 177 . 38 SER H H 9.25 0.01 1 178 . 38 SER HA H 4.89 0.01 1 179 . 38 SER HB3 H 3.99 0.01 2 180 . 39 GLY H H 8.36 0.01 1 181 . 39 GLY HA2 H 4.80 0.01 2 182 . 39 GLY HA3 H 3.90 0.01 2 183 . 40 ASP H H 9.25 0.01 1 184 . 40 ASP HA H 4.41 0.01 1 185 . 40 ASP HB3 H 3.02 0.01 2 186 . 41 ARG H H 8.05 0.01 1 187 . 41 ARG HA H 4.65 0.01 1 188 . 41 ARG HB2 H 2.33 0.01 2 189 . 41 ARG HB3 H 1.08 0.01 2 190 . 41 ARG HG2 H 1.26 0.01 2 191 . 41 ARG HG3 H 0.38 0.01 2 192 . 41 ARG HD2 H 2.92 0.01 2 193 . 41 ARG HD3 H 2.77 0.01 2 194 . 41 ARG HE H 7.64 0.01 1 195 . 41 ARG HH11 H 6.26 0.01 2 196 . 41 ARG HH12 H 6.73 0.01 2 197 . 41 ARG HH21 H 6.76 0.01 2 198 . 41 ARG HH22 H 6.19 0.01 2 199 . 42 CYS H H 7.70 0.01 1 200 . 42 CYS HA H 3.90 0.01 1 201 . 42 CYS HB2 H 3.56 0.01 2 202 . 42 CYS HB3 H 3.14 0.01 2 203 . 43 GLU H H 9.08 0.01 1 204 . 43 GLU HA H 3.98 0.01 1 205 . 43 GLU HB2 H 2.05 0.01 2 206 . 43 GLU HB3 H 1.82 0.01 2 207 . 43 GLU HG3 H 2.52 0.01 2 208 . 44 HIS H H 8.79 0.01 1 209 . 44 HIS HA H 4.96 0.01 1 210 . 44 HIS HB3 H 3.04 0.01 2 211 . 44 HIS HD2 H 7.10 0.01 1 212 . 44 HIS HE1 H 8.65 0.01 1 213 . 45 ALA H H 8.68 0.01 1 214 . 45 ALA HA H 4.29 0.01 1 215 . 45 ALA HB H 1.22 0.01 1 216 . 46 ASP H H 8.29 0.01 1 217 . 46 ASP HA H 4.51 0.01 1 218 . 46 ASP HB2 H 2.74 0.01 2 219 . 46 ASP HB3 H 2.50 0.01 2 220 . 47 LEU H H 8.11 0.01 1 221 . 47 LEU HA H 4.23 0.01 1 222 . 47 LEU HB3 H 1.54 0.01 2 223 . 47 LEU HD1 H 0.85 0.01 2 224 . 47 LEU HD2 H 0.77 0.01 2 225 . 48 LEU H H 8.09 0.01 1 226 . 48 LEU HA H 4.29 0.01 1 227 . 48 LEU HB3 H 1.53 0.01 2 228 . 48 LEU HD1 H 0.87 0.01 2 229 . 48 LEU HD2 H 0.81 0.01 2 230 . 49 ALA H H 8.23 0.01 1 231 . 49 ALA HA H 4.29 0.01 1 232 . 49 ALA HB H 1.35 0.01 1 stop_ save_