data_4410 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Three dimensional structure of Selenocosmia huwena Lectin-I (SHL-I) from the venom of the spider Selenocosmia huwena by 2D-NMR ; _BMRB_accession_number 4410 _BMRB_flat_file_name bmr4410.str _Entry_type original _Submission_date 1999-09-16 _Accession_date 1999-09-16 _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 Lu Shanyun . . 2 Liang Songping . . 3 Gu Xiaocheng . . 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" 173 "coupling constants" 11 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 1999-10-26 original author . stop_ _Original_release_date 1999-10-26 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full ; Lu, S., Liang, S., and Gu, X., "Three dimensional structure of Selenocosmia huwena Lectin-I (SHL-I) from the venom of the spider Selenocosmia huwena by 2D-NMR," J. Protein Chem. 18, 609-617 (1999). ; _Citation_title ; Three dimensional structure of Selenocosmia huwena Lectin-I (SHL-I) from the venom of the spider Selenocosmia huwena by 2D-NMR ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code 99452410 _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Lu Shanyun . . 2 Liang Songping . . 3 Gu Xiaocheng . . stop_ _Journal_abbreviation 'J. Protein Chem.' _Journal_volume 18 _Journal_issue . _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 609 _Page_last 617 _Year 1999 _Details . loop_ _Keyword lectin stop_ save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Liang SP, Pan X, "A lectin-like peptide isolated from the venom of the Chinese bird spider Selenocosmia huwena," Toxicon. 1995 Jul;33(7):875-82. ; _Citation_title 'A lectin-like peptide isolated from the venom of the Chinese bird spider Selenocosmia huwena.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 8588212 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Liang 'S P' P. . 2 Pan X . . stop_ _Journal_abbreviation Toxicon _Journal_name_full 'Toxicon : official journal of the International Society on Toxinology' _Journal_volume 33 _Journal_issue 7 _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 875 _Page_last 882 _Year 1995 _Details ; A peptide with haemagglutination activity was isolated from the venom of the Chinese bird spider Selenocosmia huwena by means of ion-exchange and reverse-phase high-performance liquid chromatography. This peptide, named SHLP-I, agglutinates human and mice erythrocytes at a minimum concentration of 125 micrograms/ml and 31 micrograms/ml, respectively. It consists of 32 amino acid residues including 3 Trp and 6 Cys residues, the latter of which form three disulfide bounds. The complete amino acid sequence was determined. The N-terminal and C-terminal residues were Gly and Trp, respectively. SHLP-I shows homology with a fragment of great nettle lectin and with huwentoxin-I from the venom of the same spider. ; save_ ################################## # Molecular system description # ################################## save_system_SHL-I _Saveframe_category molecular_system _Mol_system_name SHL-I _Abbreviation_common SHL-I _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label SHL-I $SHL-I stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'fully oxidized' loop_ _Biological_function lectin stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_SHL-I _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'Selenocosmia huwena lectin-I' _Abbreviation_common SHL-I _Molecular_mass . _Mol_thiol_state . _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 32 _Mol_residue_sequence ; GCLGDKCDYNNGCCSGYVCS RTWKWCVLAGPW ; loop_ _Residue_seq_code _Residue_label 1 GLY 2 CYS 3 LEU 4 GLY 5 ASP 6 LYS 7 CYS 8 ASP 9 TYR 10 ASN 11 ASN 12 GLY 13 CYS 14 CYS 15 SER 16 GLY 17 TYR 18 VAL 19 CYS 20 SER 21 ARG 22 THR 23 TRP 24 LYS 25 TRP 26 CYS 27 VAL 28 LEU 29 ALA 30 GLY 31 PRO 32 TRP stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-08-11 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 1QK7 "Solution Structure Of Selenocosmia Huwena Lectin-I(Shl-I) By 2d-Nmr" 100.00 32 100.00 100.00 3.03e-13 GB AAB36098 "SHLP-I=lectin-like haemagglutinating peptide [Selenocosmia huwena=Chinese bird spiders, venom, Peptide, 32 aa]" 100.00 32 100.00 100.00 3.03e-13 GB AAP33075 "huwenlectin-I precursor [Haplopelma schmidti]" 100.00 83 100.00 100.00 9.91e-15 GB ABY77717 "SHL-Ia1 precursor [Haplopelma schmidti]" 100.00 83 100.00 100.00 1.20e-14 GB ABY77719 "SHL-Ia3 precursor [Haplopelma schmidti]" 100.00 83 100.00 100.00 1.06e-14 GB ABY77720 "SHL-Ia4 precursor [Haplopelma schmidti]" 100.00 83 100.00 100.00 9.60e-15 SP B3FIS0 "RecName: Full=U5-theraphotoxin-Hs1c; Short=U5-TRTX-Hs1c; AltName: Full=Lectin SHL-1a1; AltName: Full=Lectin SHL-Ia1; Flags: Pre" 100.00 83 100.00 100.00 1.20e-14 SP B3FIS2 "RecName: Full=U5-theraphotoxin-Hs1a; Short=U5-TRTX-Hs1a; AltName: Full=Lectin SHL-Ia3; Flags: Precursor" 100.00 83 100.00 100.00 1.06e-14 SP B3FIS3 "RecName: Full=U5-theraphotoxin-Hs1a; Short=U5-TRTX-Hs1a; AltName: Full=Lectin SHL-Ia4; Flags: Precursor" 100.00 83 100.00 100.00 9.60e-15 SP B3FIS4 "RecName: Full=U5-theraphotoxin-Hs1a; Short=U5-TRTX-Hs1a; AltName: Full=Lectin SHL-Ia5; Flags: Precursor" 100.00 83 100.00 100.00 1.01e-14 SP B3FIS5 "RecName: Full=U5-theraphotoxin-Hs1a; Short=U5-TRTX-Hs1a; AltName: Full=Lectin SHL-Ia6; Flags: Precursor" 100.00 83 100.00 100.00 9.20e-15 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 $SHL-I 'Chinese bird spider' 29017 Eukaryota Metazoa Selenocosmia huwena 'venom gland' 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 $SHL-I 'natural source' . . . . . 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 $SHL-I 5.0 mM . stop_ save_ ############################ # Computer software used # ############################ save_SYBYL _Saveframe_category software _Name SYBYL _Version 6.3 _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer1 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model UnityPlus _Field_strength 600 _Details . save_ save_NMR_spectrometer2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DMX _Field_strength 500 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-1H_COSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H COSY' _Sample_label . save_ save_2D_1H-1H_DQF-COSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H DQF-COSY' _Sample_label . save_ save_2D_1H-1H_NOESY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label . save_ save_2D_1H-1H_TOCSY_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label . save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H COSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H DQF-COSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_Ex-cond_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 4.5 0.1 n/a temperature 283 1 K stop_ save_ save_Ex-cond_2 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH* 4.5 0.1 n/a temperature 292 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 TSP H 1 'methyl protons' ppm 0.00 internal direct . internal . 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 $Ex-cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name SHL-I _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 HA2 H 3.91 . 2 2 . 1 GLY HA3 H 3.78 . 2 3 . 2 CYS H H 7.79 . 1 4 . 2 CYS HA H 4.76 . 1 5 . 2 CYS HB2 H 3.50 . 2 6 . 2 CYS HB3 H 2.80 . 2 7 . 3 LEU H H 7.93 . 1 8 . 3 LEU HA H 3.93 . 1 9 . 3 LEU HB2 H 1.71 . 2 10 . 3 LEU HG H 1.46 . 1 11 . 3 LEU HD1 H 1.35 . 2 12 . 3 LEU HD2 H .97 . 2 13 . 4 GLY H H 9.35 . 1 14 . 4 GLY HA2 H 3.71 . 2 15 . 4 GLY HA3 H 2.85 . 2 16 . 5 ASP H H 7.70 . 1 17 . 5 ASP HA H 4.55 . 1 18 . 5 ASP HB2 H 2.85 . 2 19 . 5 ASP HB3 H 2.64 . 2 20 . 6 LYS H H 8.74 . 1 21 . 6 LYS HA H 4.50 . 1 22 . 6 LYS HB2 H 1.91 . 2 23 . 6 LYS HB3 H 1.74 . 2 24 . 6 LYS HG2 H 1.96 . 2 25 . 6 LYS HD2 H 1.86 . 2 26 . 6 LYS HD3 H 1.68 . 2 27 . 6 LYS HE2 H 3.14 . 2 28 . 6 LYS HZ H 7.68 . 1 29 . 7 CYS H H 8.19 . 1 30 . 7 CYS HA H 5.03 . 1 31 . 7 CYS HB2 H 3.23 . 2 32 . 7 CYS HB3 H 3.06 . 2 33 . 8 ASP H H 8.73 . 1 34 . 8 ASP HA H 4.59 . 1 35 . 8 ASP HB2 H 2.96 . 2 36 . 8 ASP HB3 H 2.88 . 2 37 . 9 TYR H H 8.31 . 1 38 . 9 TYR HA H 4.34 . 1 39 . 9 TYR HB2 H 3.06 . 2 40 . 9 TYR HB3 H 3.00 . 2 41 . 9 TYR HD1 H 7.14 . 1 42 . 9 TYR HD2 H 7.14 . 1 43 . 9 TYR HE1 H 6.82 . 1 44 . 9 TYR HE2 H 6.82 . 1 45 . 10 ASN H H 8.32 . 1 46 . 10 ASN HA H 4.59 . 1 47 . 10 ASN HB2 H 2.81 . 2 48 . 10 ASN HB3 H 2.69 . 2 49 . 10 ASN HD21 H 7.01 . 2 50 . 10 ASN HD22 H 7.76 . 2 51 . 11 ASN H H 8.09 . 1 52 . 11 ASN HA H 4.78 . 1 53 . 11 ASN HB2 H 2.74 . 2 54 . 11 ASN HB3 H 2.61 . 2 55 . 11 ASN HD21 H 6.93 . 2 56 . 11 ASN HD22 H 7.95 . 2 57 . 12 GLY H H 8.36 . 1 58 . 12 GLY HA2 H 4.26 . 2 59 . 12 GLY HA3 H 3.93 . 2 60 . 13 CYS H H 8.53 . 1 61 . 13 CYS HA H 4.92 . 1 62 . 13 CYS HB2 H 3.06 . 2 63 . 13 CYS HB3 H 2.72 . 2 64 . 14 CYS H H 7.81 . 1 65 . 14 CYS HA H 4.76 . 1 66 . 14 CYS HB2 H 3.40 . 2 67 . 14 CYS HB3 H 3.16 . 2 68 . 15 SER H H 8.43 . 1 69 . 15 SER HA H 4.31 . 1 70 . 15 SER HB2 H 3.87 . 2 71 . 16 GLY H H 9.08 . 1 72 . 16 GLY HA2 H 4.38 . 2 73 . 16 GLY HA3 H 3.67 . 2 74 . 17 TYR H H 8.54 . 1 75 . 17 TYR HA H 5.28 . 1 76 . 17 TYR HB2 H 3.23 . 2 77 . 17 TYR HB3 H 2.70 . 2 78 . 17 TYR HD1 H 6.74 . 1 79 . 17 TYR HD2 H 6.74 . 1 80 . 17 TYR HE1 H 6.97 . 1 81 . 17 TYR HE2 H 6.97 . 1 82 . 18 VAL H H 9.85 . 1 83 . 18 VAL HA H 4.62 . 1 84 . 18 VAL HB H 2.10 . 1 85 . 18 VAL HG1 H 0.94 . 2 86 . 18 VAL HG2 H 0.84 . 2 87 . 19 CYS H H 9.02 . 1 88 . 19 CYS HA H 4.72 . 1 89 . 19 CYS HB2 H 2.85 . 2 90 . 19 CYS HB3 H 2.65 . 2 91 . 20 SER H H 8.42 . 1 92 . 20 SER HA H 4.40 . 1 93 . 20 SER HB2 H 3.92 . 2 94 . 21 ARG H H 9.26 . 1 95 . 21 ARG HA H 4.06 . 1 96 . 21 ARG HB2 H 1.82 . 2 97 . 21 ARG HB3 H 1.62 . 2 98 . 21 ARG HG2 H 1.85 . 2 99 . 21 ARG HG3 H 1.72 . 2 100 . 21 ARG HD2 H 3.14 . 2 101 . 22 THR H H 7.57 . 1 102 . 22 THR HA H 3.73 . 1 103 . 22 THR HB H 3.03 . 1 104 . 22 THR HG2 H 0.16 . 1 105 . 23 TRP H H 7.25 . 1 106 . 23 TRP HA H 4.15 . 1 107 . 23 TRP HB2 H 1.33 . 2 108 . 23 TRP HB3 H 1.02 . 2 109 . 23 TRP HD1 H 6.87 . 1 110 . 23 TRP HE1 H 10.13 . 1 111 . 23 TRP HE3 H 7.70 . 1 112 . 23 TRP HZ2 H 7.48 . 1 113 . 23 TRP HZ3 H 7.16 . 1 114 . 23 TRP HH2 H 7.24 . 1 115 . 24 LYS H H 7.85 . 1 116 . 24 LYS HA H 3.87 . 1 117 . 24 LYS HB2 H 2.20 . 2 118 . 24 LYS HB3 H 2.13 . 2 119 . 24 LYS HG2 H 1.27 . 2 120 . 24 LYS HG3 H 1.22 . 2 121 . 24 LYS HD2 H 1.65 . 2 122 . 24 LYS HD3 H 1.41 . 2 123 . 24 LYS HE2 H 2.97 . 2 124 . 24 LYS HZ H 7.51 . 1 125 . 25 TRP H H 7.05 . 1 126 . 25 TRP HA H 5.61 . 1 127 . 25 TRP HB2 H 3.23 . 2 128 . 25 TRP HB3 H 2.77 . 2 129 . 25 TRP HD1 H 7.09 . 1 130 . 25 TRP HE1 H 10.21 . 1 131 . 25 TRP HE3 H 7.55 . 1 132 . 25 TRP HZ2 H 6.65 . 1 133 . 25 TRP HZ3 H 7.18 . 1 134 . 25 TRP HH2 H 6.83 . 1 135 . 26 CYS H H 9.22 . 1 136 . 26 CYS HA H 5.32 . 1 137 . 26 CYS HB2 H 3.41 . 2 138 . 26 CYS HB3 H 2.84 . 2 139 . 27 VAL H H 9.86 . 1 140 . 27 VAL HA H 5.08 . 1 141 . 27 VAL HB H 2.55 . 1 142 . 27 VAL HG1 H 1.40 . 2 143 . 27 VAL HG2 H 1.01 . 2 144 . 28 LEU H H 8.69 . 1 145 . 28 LEU HA H 3.42 . 1 146 . 28 LEU HB2 H 1.47 . 2 147 . 28 LEU HB3 H 1.41 . 2 148 . 28 LEU HG H 1.45 . 1 149 . 28 LEU HD1 H .72 . 2 150 . 28 LEU HD2 H .58 . 2 151 . 29 ALA H H 8.13 . 1 152 . 29 ALA HA H 4.18 . 1 153 . 29 ALA HB H 1.23 . 1 154 . 30 GLY H H 8.03 . 1 155 . 30 GLY HA2 H 4.02 . 2 156 . 30 GLY HA3 H 3.83 . 2 157 . 31 PRO HA H 4.37 . 1 158 . 31 PRO HB2 H 2.03 . 2 159 . 31 PRO HB3 H 1.79 . 2 160 . 31 PRO HG2 H 1.62 . 2 161 . 31 PRO HG3 H 1.86 . 2 162 . 31 PRO HD2 H 3.42 . 2 163 . 31 PRO HD3 H 3.34 . 2 164 . 32 TRP H H 7.89 . 1 165 . 32 TRP HA H 4.64 . 1 166 . 32 TRP HB2 H 3.38 . 2 167 . 32 TRP HB3 H 3.26 . 2 168 . 32 TRP HD1 H 7.25 . 1 169 . 32 TRP HE1 H 10.15 . 1 170 . 32 TRP HE3 H 7.71 . 1 171 . 32 TRP HZ2 H 7.41 . 1 172 . 32 TRP HZ3 H 7.12 . 1 173 . 32 TRP HH2 H 7.19 . 1 stop_ save_ ######################## # Coupling constants # ######################## save_J_values_set_1 _Saveframe_category coupling_constants _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $Ex-cond_1 _Spectrometer_frequency_1H 600 _Mol_system_component_name SHL-I _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 6 LYS H 6 LYS HA 3.5 . . . 2 3JHNHA 10 ASN H 10 ASN HA 9.5 . . . 3 3JHNHA 13 CYS H 13 CYS HA 8.3 . . . 4 3JHNHA 15 SER H 15 SER HA 4.9 . . . 5 3JHNHA 17 TYR H 17 TYR HA 10.4 . . . 6 3JHNHA 18 VAL H 18 VAL HA 9.2 . . . 7 3JHNHA 20 SER H 20 SER HA 5.2 . . . 8 3JHNHA 21 ARG H 21 ARG HA 3.4 . . . 9 3JHNHA 26 CYS H 26 CYS HA 2.6 . . . 10 3JHNHA 27 VAL H 27 VAL HA 12.0 . . . 11 3JHNHA 28 LEU H 28 LEU HA 5.3 . . . stop_ save_