data_5183 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; BetaCore, a designed water soluble four-stranded antiparallel b-sheet protein ; _BMRB_accession_number 5183 _BMRB_flat_file_name bmr5183.str _Entry_type original _Submission_date 2001-10-18 _Accession_date 2001-10-18 _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 Carulla Natalia . . 2 Woodward Clare . . 3 Barany George . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 2 coupling_constants 2 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 236 "15N chemical shifts" 17 "coupling constants" 17 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2008-07-16 update BMRB 'Updating non-standard residue' stop_ _Original_release_date 2001-10-18 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; BetaCore, a Designed Water Soluble Four-stranded Antiparallel Beta-sheet Protein ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code 22016650 _PubMed_ID 12021452 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Carulla Natalia . . 2 Woodward Clare . . 3 Barany George . . stop_ _Journal_abbreviation 'Protein Sci.' _Journal_volume 11 _Journal_issue 6 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 1539 _Page_last 1551 _Year 2002 _Details . save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Carulla, N.; Woodward, C.; Barany, G. Synthesis and Characterization of a b-Hairpin Peptide That Represents a 'Core Module' of Bovine Pancreatic Trypsin Inhibitor (BPTI). Biochemistry, 39, 7927-7937 (2000). ; _Citation_title ; Synthesis and characterization of a beta-hairpin peptide that represents a 'core module' of bovine pancreatic trypsin inhibitor (BPTI). ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 10891073 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Carulla N. . . 2 Woodward C. . . 3 Barany G. . . stop_ _Journal_abbreviation Biochemistry _Journal_name_full Biochemistry _Journal_volume 39 _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 7927 _Page_last 7937 _Year 2000 _Details ; A new strategy for the design and construction of peptide fragments that can achieve defined, nativelike secondary structure is presented. The strategy is based upon the hypothesis that 'core elements' of a protein, synthesized in a single polypeptide molecule, will favor nativelike structure, and that by incorporating a cross-link, nativelike core structure will dominate the ensemble as the more extended conformations are excluded. 'Core elements' are the elements of packed secondary structure that contain the slowest exchanging backbone amide protons in the native protein. The 'core elements' in bovine pancreatic trypsin inhibitor (BPTI) are the two long strands of antiparallel beta-sheet (residues 18-24 and 29-35) and the small beta-bridge (residues 43-44). To test the design strategy, we synthesized an 'oxidized core module', which contains the antiparallel strands connected by a modified reverse turn (A27 replaced by D), a natural disulfide cross-link at the open end of the hairpin, and N- and C-termini blocking groups. A peptide with identical sequence but lacking the disulfide cross-link at the open end was used as the 'reduced core module' control. The conformational behavior of both peptides was examined using (1)H NMR spectroscopy. Chemical shift dispersion, chemical shift deviation from random coil values, sequential and long-range NOEs, and H/D amide exchange rates were compared for the two peptides. We conclude that the ensemble of oxidized and reduced core module conformations samples both nativelike 4:4 and non-native 3:5 beta-hairpin structure, and that the oxidized module samples nativelike structure for a greater fraction of the time than the reduced module. ; save_ save_ref_2 _Saveframe_category citation _Citation_full ; Carulla, N.; Woodward, C.; Barany, G. Toward New Designed Proteins Derived from Bovine Pancreatic Trypsin Inhibitor (BPTI): Covalent Cross-Linking of Two 'Core Modules' by Oxime-Forming Ligation. Bioconjugate Chemistry, 12, 726-741 (2001). ; _Citation_title ; Toward new designed proteins derived from bovine pancreatic trypsin inhibitor (BPTI): covalent cross-linking of two 'core modules' by oxime-forming ligation. ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11562191 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Carulla N. . . 2 Woodward C. . . 3 Barany G. . . stop_ _Journal_abbreviation 'Bioconjug. Chem.' _Journal_name_full 'Bioconjugate chemistry' _Journal_volume 12 _Journal_issue 5 _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 726 _Page_last 741 _Year . _Details ; A 25-residue disulfide-cross-linked peptide, termed 'oxidized core module' (OxCM), that includes essentially all of the secondary structural elements of bovine pancreatic trypsin inhibitor (BPTI) most refractory to hydrogen exchange, was shown previously to favor nativelike beta-sheet structure [Carulla, N., Woodward, C., and Barany, G. (2000) Synthesis and Characterization of a beta-Hairpin Peptide That Represents a 'Core Module' of Bovine Pancreatic Trypsin Inhibitor (BPTI). Biochemistry 39, 7927-7937]. The present work prepares to explore the hypothesis that the energies of nativelike conformations, relative to other possible conformations, could be decreased further by covalent linkage of two OxCMs. Optimized syntheses of six approximately 50-residue OxCM dimers are reported herein, featuring appropriate monomer modifications followed by oxime-forming ligation chemistry to create covalent cross-links at various positions and with differing lengths. Several side reactions were recognized through this work, and modified procedures to lessen or mitigate their occurrence were developed. Particularly noteworthy, guanidine or urea denaturants that were included as peptide-solubilizing components for some reaction mixtures were proven to form adducts with glyoxylyl moieties, thus affecting rates and outcomes. All six synthetic OxCM dimers were characterized by 1D (1)H NMR; three of them showed considerable chemical shift dispersion suggestive of self-association and mutual stabilization between the monomer units. ; save_ ################################## # Molecular system description # ################################## save_system_BetaCore _Saveframe_category molecular_system _Mol_system_name BetaCore _Abbreviation_common BetaCore _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'core module I' $CM_I 'core module II' $CM_II stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state dimer _System_paramagnetic no _System_thiol_state 'all disulfide bound' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_CM_I _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common BetaCore _Abbreviation_common BetaCore _Molecular_mass . _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 26 _Mol_residue_sequence ; XKAXIIRYFYNAKDGLXQTF VYGGCX ; loop_ _Residue_seq_code _Residue_label 1 MPT 2 LYS 3 ALA 4 CLG 5 ILE 6 ILE 7 ARG 8 TYR 9 PHE 10 TYR 11 ASN 12 ALA 13 LYS 14 ASP 15 GLY 16 LEU 17 ABA 18 GLN 19 THR 20 PHE 21 VAL 22 TYR 23 GLY 24 GLY 25 CYS 26 NH2 stop_ _Sequence_homology_query_date 2008-03-24 _Sequence_homology_query_revised_last_date 2008-02-11 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 1K09 'A Chain A, Solution Structure Of Betacore,A Designed Water Soluble Four-Stranded AntiparallelB-Sheet Protein' 100.00 25 100 100 4e-04 stop_ save_ save_CM_II _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common BetaCore _Abbreviation_common BetaCore _Molecular_mass . _Mol_thiol_state 'all disulfide bound' _Details . _Residue_count 26 _Mol_residue_sequence ; XKARIIRYFYNAKDGXXQTF VYGGCX ; loop_ _Residue_seq_code _Residue_label 1 MPT 2 LYS 3 ALA 4 ARG 5 ILE 6 ILE 7 ARG 8 TYR 9 PHE 10 TYR 11 ASN 12 ALA 13 LYS 14 ASP 15 GLY 16 CLH 17 ABA 18 GLN 19 THR 20 PHE 21 VAL 22 TYR 23 GLY 24 GLY 25 CYS 26 NH2 stop_ _Sequence_homology_query_date 2008-03-24 _Sequence_homology_query_revised_last_date 2007-11-12 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 1K09 'B Chain B, Solution Structure Of Betacore,A Designed Water Soluble Four-Stranded AntiparallelB-Sheet Protein' 100.00 25 100 100 3e-04 stop_ save_ ###################### # Polymer residues # ###################### save_chem_comp_CLG _Saveframe_category polymer_residue _Mol_type 'L-peptide linking' _Name_common '2-AMINO-6-[2-(2-AMINOOXY-ACETYLAMINO)-ACETYLAMINO]-HEXANOIC ACID' _BMRB_code . _PDB_code CLG _Standard_residue_derivative . _Molecular_mass 276.290 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Thu Jul 21 13:23:42 2011 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? CA CA C . 0 . ? CB CB C . 0 . ? CG CG C . 0 . ? CD CD C . 0 . ? CE CE C . 0 . ? NZ NZ N . 0 . ? CH CH C . 0 . ? OI OI O . 0 . ? CI CI C . 0 . ? NJ NJ N . 0 . ? CK CK C . 0 . ? OL OL O . 0 . ? C C C . 0 . ? O O O . 0 . ? CL CL C . 0 . ? OM OM O . 0 . ? NM NM N . 0 . ? OXT OXT O . 0 . ? H H H . 0 . ? H2 H2 H . 0 . ? HA HA H . 0 . ? HB2 HB2 H . 0 . ? HB3 HB3 H . 0 . ? HG2 HG2 H . 0 . ? HG3 HG3 H . 0 . ? HD2 HD2 H . 0 . ? HD3 HD3 H . 0 . ? HE2 HE2 H . 0 . ? HE3 HE3 H . 0 . ? HZ HZ H . 0 . ? HI1 HI1 H . 0 . ? HI2 HI2 H . 0 . ? HNJ HNJ H . 0 . ? HL1 HL1 H . 0 . ? HL2 HL2 H . 0 . ? HNM1 HNM1 H . 0 . ? HNM2 HNM2 H . 0 . ? HXT HXT H . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name SING N CA ? ? SING N H ? ? SING N H2 ? ? SING CA CB ? ? SING CA C ? ? SING CA HA ? ? SING CB CG ? ? SING CB HB2 ? ? SING CB HB3 ? ? SING CG CD ? ? SING CG HG2 ? ? SING CG HG3 ? ? SING CD CE ? ? SING CD HD2 ? ? SING CD HD3 ? ? SING CE NZ ? ? SING CE HE2 ? ? SING CE HE3 ? ? SING NZ CH ? ? SING NZ HZ ? ? DOUB CH OI ? ? SING CH CI ? ? SING CI NJ ? ? SING CI HI1 ? ? SING CI HI2 ? ? SING NJ CK ? ? SING NJ HNJ ? ? DOUB CK OL ? ? SING CK CL ? ? DOUB C O ? ? SING C OXT ? ? SING CL OM ? ? SING CL HL1 ? ? SING CL HL2 ? ? SING OM NM ? ? SING NM HNM1 ? ? SING NM HNM2 ? ? SING OXT HXT ? ? stop_ save_ save_chem_comp_CLH _Saveframe_category polymer_residue _Mol_type 'L-peptide linking' _Name_common '2-AMINO-6-[2-(2-OXO-ACETYLAMINO)-ACETYLAMINO]-HEXANOIC ACID' _BMRB_code . _PDB_code CLH _Standard_residue_derivative . _Molecular_mass 259.259 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Thu Jul 21 13:26:03 2011 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? CA CA C . 0 . ? CB CB C . 0 . ? CG CG C . 0 . ? CD CD C . 0 . ? CE CE C . 0 . ? NZ NZ N . 0 . ? CH CH C . 0 . ? OI OI O . 0 . ? CI CI C . 0 . ? NJ NJ N . 0 . ? CK CK C . 0 . ? OL OL O . 0 . ? C C C . 0 . ? O O O . 0 . ? CL CL C . 0 . ? OM OM O . 0 . ? OXT OXT O . 0 . ? H H H . 0 . ? H2 H2 H . 0 . ? HA HA H . 0 . ? HB2 HB2 H . 0 . ? HB3 HB3 H . 0 . ? HG2 HG2 H . 0 . ? HG3 HG3 H . 0 . ? HD2 HD2 H . 0 . ? HD3 HD3 H . 0 . ? HE2 HE2 H . 0 . ? HE3 HE3 H . 0 . ? HZ HZ H . 0 . ? HI1 HI1 H . 0 . ? HI2 HI2 H . 0 . ? HNJ HNJ H . 0 . ? HL HL H . 0 . ? HXT HXT H . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name SING N CA ? ? SING N H ? ? SING N H2 ? ? SING CA CB ? ? SING CA C ? ? SING CA HA ? ? SING CB CG ? ? SING CB HB2 ? ? SING CB HB3 ? ? SING CG CD ? ? SING CG HG2 ? ? SING CG HG3 ? ? SING CD CE ? ? SING CD HD2 ? ? SING CD HD3 ? ? SING CE NZ ? ? SING CE HE2 ? ? SING CE HE3 ? ? SING NZ CH ? ? SING NZ HZ ? ? DOUB CH OI ? ? SING CH CI ? ? SING CI NJ ? ? SING CI HI1 ? ? SING CI HI2 ? ? SING NJ CK ? ? SING NJ HNJ ? ? DOUB CK OL ? ? SING CK CL ? ? DOUB C O ? ? SING C OXT ? ? DOUB CL OM ? ? SING CL HL ? ? SING OXT HXT ? ? stop_ save_ save_chem_comp_MPT _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common 'BETA-MERCAPTOPROPIONIC ACID' _BMRB_code . _PDB_code MPT _Standard_residue_derivative . _Molecular_mass 106.144 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Thu Jul 21 13:27:34 2011 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons CA CA C . 0 . ? C C C . 0 . ? O O O . 0 . ? CB CB C . 0 . ? SG SG S . 0 . ? OXT OXT O . 0 . ? HA1 HA1 H . 0 . ? HA2 HA2 H . 0 . ? HB1 HB1 H . 0 . ? HB2 HB2 H . 0 . ? HG HG H . 0 . ? HXT HXT H . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name SING CA C ? ? SING CA CB ? ? SING CA HA1 ? ? SING CA HA2 ? ? DOUB C O ? ? SING C OXT ? ? SING CB SG ? ? SING CB HB1 ? ? SING CB HB2 ? ? SING SG HG ? ? SING OXT HXT ? ? stop_ save_ save_chem_comp_ABA _Saveframe_category polymer_residue _Mol_type 'L-peptide linking' _Name_common 'ALPHA-AMINOBUTYRIC ACID' _BMRB_code . _PDB_code ABA _Standard_residue_derivative . _Molecular_mass 103.120 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Thu Jul 21 13:28:44 2011 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? CA CA C . 0 . ? C C C . 0 . ? O O O . 0 . ? CB CB C . 0 . ? CG CG C . 0 . ? OXT OXT O . 0 . ? H H H . 0 . ? HN2 HN2 H . 0 . ? HA HA H . 0 . ? HB3 HB3 H . 0 . ? HB2 HB2 H . 0 . ? HG1 HG1 H . 0 . ? HG3 HG3 H . 0 . ? HG2 HG2 H . 0 . ? HXT HXT H . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name SING N CA ? ? SING N H ? ? SING N HN2 ? ? SING CA C ? ? SING CA CB ? ? SING CA HA ? ? DOUB C O ? ? SING C OXT ? ? SING CB CG ? ? SING CB HB3 ? ? SING CB HB2 ? ? SING CG HG1 ? ? SING CG HG3 ? ? SING CG HG2 ? ? SING OXT HXT ? ? stop_ save_ save_chem_comp_NH2 _Saveframe_category polymer_residue _Mol_type non-polymer _Name_common 'AMINO GROUP' _BMRB_code . _PDB_code NH2 _Standard_residue_derivative . _Molecular_mass 16.023 _Mol_paramagnetic . _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Thu Jul 21 10:41:13 2011 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? HN1 HN1 H . 0 . ? HN2 HN2 H . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name SING N HN1 ? ? SING N HN2 ? ? stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $CM_I Bull 9913 Eukaryota Metazoa Bos taurus $CM_II Bull 9913 Eukaryota Metazoa Bos taurus 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 $CM_I 'chemical synthesis' . . . . . $CM_II 'chemical synthesis' . . . . . 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 $CM_I 0.4 mM '[38% selectively 15N]' $CM_II 0.4 mM '[38% selectively 15N]' stop_ save_ ############################ # Computer software used # ############################ save_NMRPipe _Saveframe_category software _Name NMRPipe _Version . _Details . save_ save_NMRView _Saveframe_category software _Name NMRView _Version . _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model Inova _Field_strength 600 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model Inova _Field_strength 800 _Details . save_ ############################# # NMR applied experiments # ############################# save_1H-1H_NOESY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H NOESY' _Sample_label $sample_1 save_ save_1H-1H_TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1H-1H TOCSY' _Sample_label $sample_1 save_ save_15N-1H_HSQC_3 _Saveframe_category NMR_applied_experiment _Experiment_name '15N-1H HSQC' _Sample_label $sample_1 save_ save_15N-1H_HSQC-TOCSY_4 _Saveframe_category NMR_applied_experiment _Experiment_name '15N-1H HSQC-TOCSY' _Sample_label $sample_1 save_ save_15N-1H_HSQC-NOESY_5 _Saveframe_category NMR_applied_experiment _Experiment_name '15N-1H HSQC-NOESY' _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_cond_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 3 0.2 n/a temperature 288 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 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_shifts_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '1H-1H NOESY' '1H-1H TOCSY' '15N-1H HSQC' '15N-1H HSQC-TOCSY' '15N-1H HSQC-NOESY' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'core module 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 . 2 LYS H H 8.44 0.02 1 2 . 2 LYS HA H 4.19 0.02 1 3 . 2 LYS HB2 H 1.87 0.02 2 4 . 2 LYS HB3 H 1.79 0.02 2 5 . 2 LYS HG2 H 1.49 0.02 1 6 . 2 LYS HG3 H 1.49 0.02 1 7 . 2 LYS HD2 H 1.69 0.02 1 8 . 2 LYS HD3 H 1.69 0.02 1 9 . 2 LYS HE2 H 2.99 0.02 1 10 . 2 LYS HE3 H 2.99 0.02 1 11 . 3 ALA H H 8.00 0.02 1 12 . 3 ALA HA H 4.34 0.02 1 13 . 3 ALA HB H 1.39 0.02 1 14 . 3 ALA N N 122.01 0.02 1 15 . 4 CLG H H 7.76 0.02 1 16 . 4 CLG HA H 4.68 0.02 1 17 . 4 CLG HB2 H 2.07 0.02 2 18 . 4 CLG HB3 H 1.95 0.02 2 19 . 4 CLG HG2 H 1.47 0.02 1 20 . 4 CLG HG3 H 1.47 0.02 1 21 . 4 CLG HD2 H 1.67 0.02 1 22 . 4 CLG HD3 H 1.67 0.02 1 23 . 4 CLG HE2 H 2.30 0.02 2 24 . 4 CLG HE3 H 2.21 0.02 2 25 . 5 ILE H H 8.68 0.02 1 26 . 5 ILE HA H 4.38 0.02 1 27 . 5 ILE HB H 1.63 0.02 1 28 . 5 ILE HD1 H 0.548 0.02 1 29 . 7 ARG H H 9.34 0.02 1 30 . 7 ARG HA H 4.75 0.02 1 31 . 8 TYR H H 8.13 0.02 1 32 . 8 TYR HA H 5.93 0.02 1 33 . 8 TYR HB2 H 3.13 0.02 2 34 . 8 TYR HB3 H 2.85 0.02 2 35 . 8 TYR HD1 H 6.97 0.02 1 36 . 8 TYR HD2 H 6.97 0.02 1 37 . 8 TYR HE1 H 6.75 0.02 1 38 . 8 TYR HE2 H 6.75 0.02 1 39 . 9 PHE H H 9.28 0.02 1 40 . 9 PHE HA H 4.93 0.02 1 41 . 9 PHE HB2 H 3.30 0.02 2 42 . 9 PHE HB3 H 3.04 0.02 2 43 . 9 PHE HD1 H 7.27 0.02 1 44 . 9 PHE HD2 H 7.27 0.02 1 45 . 9 PHE HE1 H 7.22 0.02 1 46 . 9 PHE HE2 H 7.22 0.02 1 47 . 9 PHE N N 118.41 0.02 1 48 . 10 TYR H H 8.91 0.02 1 49 . 10 TYR HA H 5.11 0.02 1 50 . 10 TYR HB2 H 2.30 0.02 2 51 . 10 TYR HB3 H 2.06 0.02 2 52 . 10 TYR HD1 H 6.59 0.02 1 53 . 10 TYR HD2 H 6.59 0.02 1 54 . 10 TYR HE1 H 6.42 0.02 1 55 . 10 TYR HE2 H 6.42 0.02 1 56 . 11 ASN H H 8.09 0.02 1 57 . 11 ASN HA H 4.65 0.02 1 58 . 11 ASN HB2 H 3.02 0.02 2 59 . 11 ASN HB3 H 2.66 0.02 2 60 . 12 ALA H H 8.5 0.02 1 61 . 12 ALA HA H 3.76 0.02 1 62 . 12 ALA HB H 1.53 0.02 1 63 . 12 ALA N N 126.68 0.02 1 64 . 13 LYS H H 8.14 0.02 1 65 . 13 LYS HA H 4.05 0.02 1 66 . 14 ASP H H 7.27 0.02 1 67 . 14 ASP HA H 4.71 0.02 1 68 . 14 ASP HB2 H 2.68 0.02 2 69 . 14 ASP HB3 H 2.35 0.02 2 70 . 15 GLY H H 7.86 0.02 1 71 . 15 GLY HA2 H 3.50 0.02 2 72 . 15 GLY HA3 H 3.27 0.02 2 73 . 15 GLY N N 110.53 0.02 1 74 . 16 LEU H H 6.96 0.02 1 75 . 16 LEU HA H 5.15 0.02 1 76 . 16 LEU HB2 H 1.5 0.02 1 77 . 16 LEU HB3 H 1.5 0.02 1 78 . 16 LEU HG H 1.18 0.02 1 79 . 16 LEU HD1 H 0.79 0.02 2 80 . 16 LEU HD2 H 0.59 0.02 2 81 . 16 LEU N N 115.9 0.02 1 82 . 17 ABA H H 9.38 0.02 1 83 . 17 ABA HA H 5.24 0.02 1 84 . 17 ABA HB2 H 2.23 0.02 1 85 . 17 ABA HG2 H 1.05 0.02 1 86 . 18 GLN H H 9.29 0.02 1 87 . 18 GLN HA H 5.61 0.02 1 88 . 18 GLN HB2 H 2.04 0.02 2 89 . 18 GLN HB3 H 1.90 0.02 2 90 . 18 GLN HG2 H 2.29 0.02 1 91 . 18 GLN HG3 H 2.29 0.02 1 92 . 19 THR H H 8.99 0.02 1 93 . 19 THR HA H 5.43 0.02 1 94 . 19 THR HB H 4.23 0.02 1 95 . 19 THR HG2 H 1.36 0.02 1 96 . 20 PHE H H 9.77 0.02 1 97 . 20 PHE HA H 5.1 0.02 1 98 . 20 PHE HB2 H 3.03 0.02 2 99 . 20 PHE HB3 H 2.56 0.02 2 100 . 20 PHE HD1 H 7.16 0.02 1 101 . 20 PHE HD2 H 7.16 0.02 1 102 . 20 PHE HE1 H 6.68 0.02 1 103 . 20 PHE HE2 H 6.68 0.02 1 104 . 20 PHE N N 126.38 0.02 1 105 . 21 VAL H H 9.34 0.02 1 106 . 21 VAL HA H 4.75 0.02 1 107 . 21 VAL HB H 2.05 0.02 1 108 . 21 VAL HG1 H 0.88 0.02 1 109 . 21 VAL HG2 H 0.88 0.02 1 110 . 21 VAL N N 123.28 0.02 1 111 . 22 TYR H H 9.57 0.02 1 112 . 22 TYR HA H 4.97 0.02 1 113 . 22 TYR HB2 H 3.16 0.02 2 114 . 22 TYR HB3 H 3.04 0.02 2 115 . 22 TYR HE1 H 7.18 0.02 1 116 . 22 TYR HE2 H 7.18 0.02 1 117 . 23 GLY H H 8.82 0.02 1 118 . 23 GLY HA2 H 4.31 0.02 2 119 . 23 GLY HA3 H 3.98 0.02 2 120 . 23 GLY N N 110.47 0.02 1 121 . 24 GLY H H 8.07 0.02 1 122 . 24 GLY HA2 H 3.97 0.02 1 123 . 24 GLY HA3 H 3.97 0.02 1 124 . 24 GLY N N 106.23 0.02 1 125 . 25 CYS H H 8.49 0.02 1 126 . 25 CYS HA H 4.65 0.02 1 127 . 25 CYS HB2 H 3.23 0.02 2 128 . 25 CYS HB3 H 2.98 0.02 2 stop_ save_ save_chemical_shifts_2 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '1H-1H NOESY' '1H-1H TOCSY' '15N-1H HSQC' '15N-1H HSQC-TOCSY' '15N-1H HSQC-NOESY' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'core module II' _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 . 2 LYS H H 8.36 0.02 1 2 . 2 LYS HA H 4.19 0.02 1 3 . 3 ALA H H 8.12 0.02 1 4 . 3 ALA HA H 4.34 0.02 1 5 . 3 ALA HB H 1.41 0.02 1 6 . 3 ALA N N 122.37 0.02 1 7 . 4 ARG H H 7.93 0.02 1 8 . 4 ARG HA H 4.62 0.02 1 9 . 4 ARG HB2 H 1.69 0.02 1 10 . 4 ARG HB3 H 1.69 0.02 1 11 . 4 ARG HG2 H 1.59 0.02 2 12 . 4 ARG HG3 H 1.50 0.02 2 13 . 5 ILE H H 8.68 0.02 1 14 . 5 ILE HA H 4.75 0.02 1 15 . 5 ILE HB H 1.78 0.02 1 16 . 5 ILE HG12 H 1.54 0.02 2 17 . 5 ILE HG13 H 0.96 0.02 2 18 . 5 ILE HG2 H 0.65 0.02 1 19 . 6 ILE H H 8.64 0.02 1 20 . 6 ILE HA H 4.7 0.02 1 21 . 6 ILE HG12 H 1.41 0.02 2 22 . 6 ILE HG13 H 0.97 0.02 2 23 . 6 ILE HG2 H 0.36 0.02 1 24 . 7 ARG H H 9.06 0.02 1 25 . 7 ARG HA H 5.33 0.02 1 26 . 7 ARG HB2 H 1.72 0.02 1 27 . 7 ARG HB3 H 1.72 0.02 1 28 . 7 ARG HG2 H 1.44 0.02 1 29 . 7 ARG HG3 H 1.44 0.02 1 30 . 8 TYR H H 8.79 0.02 1 31 . 8 TYR HA H 5.58 0.02 1 32 . 8 TYR HB2 H 3.15 0.02 2 33 . 8 TYR HB3 H 2.90 0.02 2 34 . 8 TYR HD1 H 6.94 0.02 1 35 . 8 TYR HD2 H 6.94 0.02 1 36 . 8 TYR HE1 H 6.51 0.02 1 37 . 8 TYR HE2 H 6.51 0.02 1 38 . 9 PHE H H 9.7 0.02 1 39 . 9 PHE HA H 5.28 0.02 1 40 . 9 PHE HB2 H 3.36 0.02 2 41 . 9 PHE HB3 H 2.97 0.02 2 42 . 9 PHE HD1 H 7.22 0.02 1 43 . 9 PHE HD2 H 7.22 0.02 1 44 . 9 PHE HE1 H 7.12 0.02 1 45 . 9 PHE HE2 H 7.12 0.02 1 46 . 9 PHE N N 126.63 0.02 1 47 . 10 TYR H H 9.28 0.02 1 48 . 10 TYR HA H 4.92 0.02 1 49 . 10 TYR HB2 H 3.30 0.02 2 50 . 10 TYR HB3 H 2.80 0.02 2 51 . 10 TYR HD1 H 6.75 0.02 1 52 . 10 TYR HD2 H 6.75 0.02 1 53 . 10 TYR HE1 H 6.42 0.02 1 54 . 10 TYR HE2 H 6.42 0.02 1 55 . 11 ASN H H 8.23 0.02 1 56 . 11 ASN HA H 4.49 0.02 1 57 . 11 ASN HB2 H 3.21 0.02 1 58 . 11 ASN HB3 H 3.21 0.02 1 59 . 12 ALA H H 8.42 0.02 1 60 . 12 ALA HA H 3.56 0.02 1 61 . 12 ALA HB H 1.3 0.02 1 62 . 12 ALA N N 124.64 0.02 1 63 . 13 LYS H H 7.95 0.02 1 64 . 13 LYS HA H 4.07 0.02 1 65 . 13 LYS HB2 H 1.87 0.02 2 66 . 13 LYS HB3 H 1.82 0.02 2 67 . 13 LYS HD2 H 1.69 0.02 1 68 . 13 LYS HD3 H 1.69 0.02 1 69 . 14 ASP H H 6.62 0.02 1 70 . 14 ASP HA H 4.65 0.02 1 71 . 15 GLY H H 7.44 0.02 1 72 . 15 GLY HA2 H 3.47 0.02 1 73 . 15 GLY HA3 H 3.47 0.02 1 74 . 15 GLY N N 108.01 0.02 1 75 . 16 CLH H H 6.78 0.02 1 76 . 16 CLH HA H 4.32 0.02 1 77 . 16 CLH HE2 H 1.56 0.02 1 78 . 16 CLH HE3 H 1.56 0.02 1 79 . 17 ABA H H 8.94 0.02 1 80 . 17 ABA HA H 5.1 0.02 1 81 . 17 ABA HB2 H 1.97 0.02 1 82 . 17 ABA HB3 H 1.97 0.02 1 83 . 17 ABA HG2 H 1.02 0.02 1 84 . 17 ABA HG3 H 1.02 0.02 1 85 . 18 GLN H H 9.34 0.02 1 86 . 18 GLN HA H 4.75 0.02 1 87 . 18 GLN HB2 H 1.95 0.02 1 88 . 18 GLN HB3 H 1.95 0.02 1 89 . 18 GLN HG2 H 2.05 0.02 1 90 . 18 GLN HG3 H 2.05 0.02 1 91 . 19 THR H H 8.55 0.02 1 92 . 19 THR HA H 5.19 0.02 1 93 . 19 THR HB H 3.85 0.02 1 94 . 19 THR HG2 H 1.06 0.02 1 95 . 20 PHE H H 9.34 0.02 1 96 . 20 PHE HA H 4.75 0.02 1 97 . 20 PHE HB2 H 2.76 0.02 1 98 . 20 PHE HB3 H 2.76 0.02 1 99 . 20 PHE N N 126.75 0.02 1 100 . 21 VAL H H 8.38 0.02 1 101 . 21 VAL HA H 4.64 0.02 1 102 . 21 VAL HB H 1.87 0.02 1 103 . 21 VAL HG1 H 0.83 0.02 1 104 . 21 VAL HG2 H 0.83 0.02 1 105 . 21 VAL N N 121.88 0.02 1 106 . 22 TYR H H 9.16 0.02 1 107 . 22 TYR HA H 4.88 0.02 1 108 . 22 TYR HB2 H 3.15 0.02 2 109 . 22 TYR HB3 H 2.91 0.02 2 110 . 22 TYR HD1 H 7.07 0.02 1 111 . 22 TYR HD2 H 7.07 0.02 1 112 . 22 TYR HE1 H 6.69 0.02 1 113 . 22 TYR HE2 H 6.69 0.02 1 114 . 23 GLY H H 8.67 0.02 1 115 . 23 GLY HA2 H 4.23 0.02 2 116 . 23 GLY HA3 H 3.80 0.02 2 117 . 23 GLY N N 110.18 0.02 1 118 . 24 GLY H H 7.99 0.02 1 119 . 24 GLY HA2 H 3.97 0.02 1 120 . 24 GLY HA3 H 3.97 0.02 1 121 . 24 GLY N N 107.17 0.02 1 122 . 25 CYS H H 8.56 0.02 1 123 . 25 CYS HA H 4.69 0.02 1 124 . 25 CYS HB2 H 3.27 0.02 1 125 . 25 CYS HB3 H 3.27 0.02 1 stop_ save_ ######################## # Coupling constants # ######################## save_coupling_constants_1 _Saveframe_category coupling_constants _Details . loop_ _Experiment_label '15N-1H HSQC' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $cond_1 _Spectrometer_frequency_1H 600 _Mol_system_component_name 'core module 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 3 ALA H 3 ALA HA 5.8 . . 1.0 2 3JHNHA 9 PHE H 9 PHE HA 8.0 . . 1.0 3 3JHNHA 12 ALA H 12 ALA HA 4.4 . . 1.0 4 3JHNHA 15 GLY H 15 GLY HA2 5.2 . . 1.0 5 3JHNHA 15 GLY H 15 GLY HA3 4.7 . . 1.0 6 3JHNHA 16 LEU H 16 LEU HA 8.5 . . 1.0 7 3JHNHA 20 PHE H 20 PHE HA 8.5 . . 1.0 8 3JHNHA 21 VAL H 21 VAL HA 8.5 . . 1.0 9 3JHNHA 23 GLY H 23 GLY HA2 4.8 . . 1.0 10 3JHNHA 23 GLY H 23 GLY HA3 4.7 . . 1.0 stop_ save_ save_coupling_constants_2 _Saveframe_category coupling_constants _Details . loop_ _Experiment_label '15N-1H HSQC' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $cond_1 _Spectrometer_frequency_1H 600 _Mol_system_component_name 'core module II' _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 ALA H 3 ALA HA 5.9 . . 1.0 2 3JHNHA 9 PHE H 9 PHE HA 8.6 . . 1.0 3 3JHNHA 12 ALA H 12 ALA HA 5.1 . . 1.0 4 3JHNHA 20 PHE H 20 PHE HA 9.2 . . 1.0 5 3JHNHA 21 VAL H 21 VAL HA 8.0 . . 1.0 6 3JHNHA 23 GLY H 23 GLY HA2 5.2 . . 1.0 7 3JHNHA 23 GLY H 23 GLY HA3 4.6 . . 1.0 stop_ save_