data_5757 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 13C, 15N solid state NMR chemical shift assignments for the microcrystallin Crh domain swapped dimer ; _BMRB_accession_number 5757 _BMRB_flat_file_name bmr5757.str _Entry_type original _Submission_date 2003-03-27 _Accession_date 2003-03-27 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details ; Chemical shifts for the monomeric (BMRB-4972) and dimeric forms of Crh show important differences due to conformational changes induced by the N-terminal 3D domain swap observed in the dimeric form ; loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Bockmann Anja . . 2 Lange Adam . . 3 Galinier Anne . . 4 Luca Sorin . . 5 Giraud Nicolas . . 6 Juy Michel . . 7 Heise Henrike . . 8 Montserret Roland . . 9 Penin Francois . . 10 Baldus Marc . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 2 stop_ loop_ _Data_type _Data_type_count "13C chemical shifts" 360 "15N chemical shifts" 90 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2003-10-16 original author . stop_ loop_ _Related_BMRB_accession_number _Relationship 4972 'Crh monomer in solution state' stop_ _Original_release_date 2003-10-16 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Solid state NMR sequential resonance assignments and conformational analysis of the 2x10.4 kDa dimeric form of the Bacillus subtilis protein Crh ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Bockmann Anja . . 2 Lange Adam . . 3 Galinier Anne . . 4 Luca Sorin . . 5 Giraud Nicolas . . 6 Juy Michel . . 7 Heise Henrike . . 8 Montserret Roland . . 9 Penin Francois . . 10 Baldus Marc . . stop_ _Journal_abbreviation 'J. Biomol. NMR' _Journal_volume 27 _Journal_issue 4 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 323 _Page_last 339 _Year 2003 _Details . loop_ _Keyword Assignments 'catabolite repression histidine-containing phosphocarrier protein (Crh)' MAS 'protein dynamics' 'protein structure' 'solid state NMR spectroscopy' stop_ save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full . _Citation_title 'Evidence for a dimerisation state of the Bacillus subtilis catabolite repression HPr-like protein, Crh.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11361074 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Penin F. . . 2 Favier A. . . 3 Montserret R. . . 4 Brutscher B. . . 5 Deutscher J. . . 6 Marion D. . . 7 Galinier D. . . stop_ _Journal_abbreviation 'J. Mol. Microbiol. Biotechnol.' _Journal_name_full 'Journal of molecular microbiology and biotechnology' _Journal_volume 3 _Journal_issue 3 _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 429 _Page_last 432 _Year 2001 _Details ; The Bacillus subtilis catabolite repression HPr (Crh) exhibits 45% sequence identity when compared to histidine-containing protein (HPr), a phosphocarrier protein of the phosphoenolpyruvate:carbohydrate phosphotransferase system. We report here that Crh preparations contain a mixture of monomers and homodimers, whereas HPr is known to be monomeric in solution. The dissociation rate of dimers is very slow (t1/2 of about 10 hours), and the percentage of dimers in Crh preparations increases with rising temperature or protein concentration. However, at temperatures above 25 degrees C and a protein concentration of 10 mg/ml, Crh dimers slowly aggregate. Typically, NMR spectra recorded at 25 degrees C showed the coexistence of both forms of Crh, while in Crh solutions kept at 35 degrees C, almost exclusively Crh monomers could be detected. Circular dichroism analysis revealed that the monomeric and dimeric forms of Crh are well folded and exhibit the same overall structure. The physiological significance of the slow Crh monomer/dimer equilibrium remains enigmatic. ; save_ save_ref_2 _Saveframe_category citation _Citation_full . _Citation_title 'Solution structure and dynamics of Crh, the Bacillus subtilis catabolite repression HPr.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11916384 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Favier Adrien . . 2 Brutscher Bernhard . . 3 Blackledge Martin . . 4 Galinier Anne . . 5 Deutscher Josef . . 6 Penin Francois . . 7 Marion Dominique . . stop_ _Journal_abbreviation 'J. Mol. Biol.' _Journal_name_full 'Journal of molecular biology' _Journal_volume 317 _Journal_issue 1 _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first 131 _Page_last 144 _Year 2002 _Details ; The solution structure and dynamics of the Bacillus subtilis HPr-like protein, Crh, have been investigated using NMR spectroscopy. Crh exhibits high sequence identity (45 %) to the histidine-containing protein (HPr), a phospho-carrier protein of the phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system, but contains no catalytic His15, the site of PEP-dependent phosphorylation in HPr. Crh also forms a mixture of monomers and dimers in solution whereas HPr is known to be monomeric. Complete backbone and side-chain assignments were obtained for the monomeric form, and 60 % of the dimer backbone resonances; allowing the identification of the Crh dimer interface from chemical-shift mapping. The conformation of Crh was determined to a precision of 0.46(+/-0.06) A for the backbone atoms, and 1.01(+/-0.08) A for the heavy atoms. The monomer structure is similar to that of known HPr 2.67(+/-0.22) A (C(alpha) rmsd), but has a few notable differences, including a change in the orientation of one of the helices (B), and a two-residue shift in beta-sheet pairing of the N-terminal strand with the beta4 strand. This shift results in a shortening of the surface loop present in HPr and consequently provides a flatter surface in the region of dimerisation contact, which may be related to the different oligomeric nature of these two proteins. A binding site of phospho-serine(P-Ser)-Crh with catabolite control protein A (CcpA) is proposed on the basis of highly conserved surface side-chains between Crh and HPr. This binding site is consistent with the model of a dimer-dimer interaction between P-Ser-Crh and CcpA. (15)N relaxation measured in the monomeric form also identified differential local mobility in the helix B which is located in the vicinity of this site. ; save_ save_ref_3 _Saveframe_category citation _Citation_full ; Dimerization of Crh by reversible 3D domain swapping induces structural adjustments to its monomeric homologue HPr Juy, M., Penin, F., Favier, A., Galinier, A., Montserret, R., Haser, R., Deutscher, J. and Bockmann, A. (2003) submitted ; _Citation_title 'Dimerization of Crh by reversible 3D domain swapping induces structural adjustments to its monomeric homologue Hpr.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 12972249 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Juy Michel . . 2 Penin Francois . . 3 Favier Adrien . . 4 Galinier Anne . . 5 Montserret Roland . . 6 Haser Richard . . 7 Deutscher Josef . . 8 Bockmann Anja . . stop_ _Journal_abbreviation 'J. Mol. Biol.' _Journal_name_full 'Journal of molecular biology' _Journal_volume 332 _Journal_issue 4 _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 767 _Page_last 776 _Year 2003 _Details ; The crystal structure of the regulatory protein Crh from Bacillus subtilis was solved at 1.8A resolution and showed an intertwined dimer formed by N-terminal beta1-strand swapping of two monomers. Comparison with the monomeric NMR structure of Crh revealed a domain swap induced conformational rearrangement of the putative interaction site with the repressor CcpA. The resulting conformation closely resembles that observed for the monomeric Crh homologue HPr, indicating that the Crh dimer is the active form binding to CcpA. An analogous dimer of HPr can be constructed without domain swapping, suggesting that HPr may dimerize upon binding to CcpA. Our data suggest that reversible 3D domain swapping of Crh might be an efficient regulatory mechanism to modulate its activity. ; save_ ################################## # Molecular system description # ################################## save_system_Crh _Saveframe_category molecular_system _Mol_system_name 'Crh dimer' _Abbreviation_common Crh _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'Crh subunit 1' $Crh 'Crh subunit 2' $Crh stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state dimer _System_paramagnetic no _System_thiol_state 'not present' loop_ _Magnetic_equivalence_ID _Magnetically_equivalent_system_component 1 'Crh subunit 1' 1 'Crh subunit 2' stop_ loop_ _Biological_function 'phospho-carrier protein' stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_Crh _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'catabolite repression HPr' _Abbreviation_common Crh _Molecular_mass 10391 _Mol_thiol_state 'not present' _Details ; Crh has been shown to exist in a slow monomer-dimer equilibrium in solution. The structure of the monomeric form could be solved by liquid state NMR spectroscopy. We recently solved the domain swapped dimer structure by X-ray crystallograpy. The microcrystall in solid Crh dimer serves us as model molecule for the development of solid state NMR methods ; ############################## # Polymer residue sequence # ############################## _Residue_count 93 _Mol_residue_sequence ; MVQQKVEVRLKTGLQARPAA LFVQEANRFTSDVFLEKDGK KVNAKSIMGLMSLAVSTGTE VTLIAQGEDEQEALEKLAAY VQEEVLQHHHHHH ; loop_ _Residue_seq_code _Residue_label 1 MET 2 VAL 3 GLN 4 GLN 5 LYS 6 VAL 7 GLU 8 VAL 9 ARG 10 LEU 11 LYS 12 THR 13 GLY 14 LEU 15 GLN 16 ALA 17 ARG 18 PRO 19 ALA 20 ALA 21 LEU 22 PHE 23 VAL 24 GLN 25 GLU 26 ALA 27 ASN 28 ARG 29 PHE 30 THR 31 SER 32 ASP 33 VAL 34 PHE 35 LEU 36 GLU 37 LYS 38 ASP 39 GLY 40 LYS 41 LYS 42 VAL 43 ASN 44 ALA 45 LYS 46 SER 47 ILE 48 MET 49 GLY 50 LEU 51 MET 52 SER 53 LEU 54 ALA 55 VAL 56 SER 57 THR 58 GLY 59 THR 60 GLU 61 VAL 62 THR 63 LEU 64 ILE 65 ALA 66 GLN 67 GLY 68 GLU 69 ASP 70 GLU 71 GLN 72 GLU 73 ALA 74 LEU 75 GLU 76 LYS 77 LEU 78 ALA 79 ALA 80 TYR 81 VAL 82 GLN 83 GLU 84 GLU 85 VAL 86 LEU 87 GLN 88 HIS 89 HIS 90 HIS 91 HIS 92 HIS 93 HIS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-01-28 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value BMRB 4972 crh_monomer 93.55 87 100.00 100.00 1.13e-53 PDB 1K1C "Solution Structure Of Crh, The Bacillus Subtilis Catabolite Repression Hpr" 90.32 84 100.00 100.00 2.37e-51 PDB 1MO1 "Crystal Structure At 1.8 Angstroms Of Seleno Methionyled Crh, The Bacillus Subtilis Catabolite Repression Containing Protein Cr" 93.55 87 97.70 97.70 1.47e-51 PDB 1MU4 "Crystal Structure At 1.8 Angstroms Of The Bacillus Subtilis Catabolite Repression Histidine Containing Protein (crh)" 93.55 87 100.00 100.00 1.13e-53 PDB 2AK7 "Structure Of A Dimeric P-Ser-Crh" 92.47 86 98.84 98.84 4.25e-52 PDB 2RLZ "Solid-State Mas Nmr Structure Of The Dimer Crh" 91.40 85 100.00 100.00 2.50e-52 DBJ BAI87096 "phosphocarrier protein Chr [Bacillus subtilis subsp. natto BEST195]" 91.40 85 100.00 100.00 2.50e-52 DBJ BAM55550 "phosphocarrier protein Chr [Bacillus subtilis BEST7613]" 91.40 85 100.00 100.00 2.50e-52 DBJ BAM59563 "phosphocarrier protein Chr [Bacillus subtilis BEST7003]" 91.40 85 100.00 100.00 2.50e-52 DBJ GAK79281 "phosphocarrier protein Chr [Bacillus subtilis Miyagi-4]" 91.40 85 100.00 100.00 2.50e-52 EMBL CAB08060 "hypothetical protein [Bacillus subtilis]" 91.40 85 100.00 100.00 2.50e-52 EMBL CAB15479 "catabolite repression HPr-like protein [Bacillus subtilis subsp. subtilis str. 168]" 91.40 85 100.00 100.00 2.50e-52 EMBL CCU60557 "Catabolite repression HPr-like protein Crh [Bacillus subtilis E1]" 91.40 85 100.00 100.00 2.50e-52 EMBL CEI59278 "HPr-like protein Crh [Bacillus subtilis]" 91.40 85 100.00 100.00 2.50e-52 EMBL CEJ79135 "HPr-like protein Crh [Bacillus sp.]" 91.40 85 100.00 100.00 2.50e-52 GB ADM39440 "catabolite repression HPr-like protein [Bacillus subtilis subsp. spizizenii str. W23]" 91.40 85 100.00 100.00 2.50e-52 GB ADV94285 "phosphocarrier protein Chr [Bacillus subtilis BSn5]" 91.40 85 100.00 100.00 2.50e-52 GB AEP88381 "HPr like protein, Crh [Bacillus subtilis subsp. spizizenii TU-B-10]" 91.40 85 100.00 100.00 2.50e-52 GB AEP92509 "HPr like protein, Crh [Bacillus subtilis subsp. subtilis str. RO-NN-1]" 91.40 85 100.00 100.00 2.50e-52 GB AFI30049 "phosphocarrier protein Chr [Bacillus sp. JS]" 91.40 85 100.00 100.00 2.50e-52 REF NP_391354 "HPr-like protein Crh [Bacillus subtilis subsp. subtilis str. 168]" 91.40 85 100.00 100.00 2.50e-52 REF WP_003219835 "MULTISPECIES: phosphate ABC transporter permease [Bacillales]" 91.40 85 100.00 100.00 2.50e-52 REF WP_024122965 "phosphate ABC transporter permease [Bacillus mojavensis]" 91.40 85 97.65 100.00 2.21e-51 REF WP_024713407 "phosphocarrier protein Chr [Bacillus tequilensis]" 91.40 85 98.82 98.82 1.38e-51 REF YP_003867749 "catabolite repression HPr-like protein [Bacillus subtilis subsp. spizizenii str. W23]" 91.40 85 100.00 100.00 2.50e-52 SP O06976 "RecName: Full=HPr-like protein Crh; AltName: Full=Catabolite repression HPr [Bacillus subtilis subsp. subtilis str. 168]" 91.40 85 100.00 100.00 2.50e-52 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $Crh 'Bacillus subtilis' 1423 Eubacteria . Bacillus subtilis 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 $Crh 'recombinant technology' . . . . . stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solid _Details 'Microcrystallin solid protein obtained by slow precipitation with PEG' loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $Crh . mg 4 20 '[U-98% 13C; U-98% 15N]' stop_ save_ ############################ # Computer software used # ############################ save_NMRPipe _Saveframe_category software _Name NMRPipe _Version 2.0 loop_ _Task 'data processing' stop_ _Details . save_ save_NMRVIEW _Saveframe_category software _Name NMRVIEW _Version 5.0 loop_ _Task assignments stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer BRUKER _Model DSX _Field_strength 500 _Details . save_ save_spectrometer _Saveframe_category NMR_spectrometer _Manufacturer BRUKER _Model DSX _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_13C-13C_proton_driven_spin_diffusion_(PDSD)_1 _Saveframe_category NMR_applied_experiment _Experiment_name '13C-13C proton driven spin diffusion (PDSD)' _Sample_label $sample_1 save_ save_13C-13C_CO_selective_J-decoupled_PDSD_2 _Saveframe_category NMR_applied_experiment _Experiment_name '13C-13C CO selective J-decoupled PDSD' _Sample_label $sample_1 save_ save_13C-13C_RFDR_3 _Saveframe_category NMR_applied_experiment _Experiment_name '13C-13C RFDR' _Sample_label $sample_1 save_ save_13C-13C_DQ_SPC-5_4 _Saveframe_category NMR_applied_experiment _Experiment_name '13C-13C DQ SPC-5' _Sample_label $sample_1 save_ save_15N-13C_NCACO_5 _Saveframe_category NMR_applied_experiment _Experiment_name '15N-13C NCACO' _Sample_label $sample_1 save_ save_15N-13C_NCOCACB_6 _Saveframe_category NMR_applied_experiment _Experiment_name '15N-13C NCOCACB' _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_Ex-cond_1 _Saveframe_category sample_conditions _Details ; The actual sample temperature is around 278K, which is not read out probe T. MAS speed is around 10 kHz. ; loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 6.9 0.2 n/a 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 H 1 'methyl protons' ppm 0 external direct . external_to_the_sample . . DSS N 15 'methyl protons' ppm 0.0 . indirect . . . 0.101329118 DSS C 13 'methyl protons' ppm 0.0 . indirect . . . 0.251449530 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 'Crh subunit 1' _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 VAL N N 116.5 0.1 1 2 . 2 VAL CA C 58.0 0.1 1 3 . 2 VAL CB C 35.1 0.1 1 4 . 2 VAL CG1 C 21.5 0.1 1 5 . 2 VAL CG2 C 21.5 0.1 1 6 . 2 VAL C C 174.0 0.1 1 7 . 3 GLN N N 82.6 0.1 1 8 . 3 GLN CA C 54.9 0.1 1 9 . 3 GLN CB C 32.7 0.1 1 10 . 3 GLN CG C 30.9 0.1 1 11 . 3 GLN C C 172.2 0.1 1 12 . 4 GLN N N 125.5 0.1 1 13 . 4 GLN CA C 55.5 0.1 1 14 . 4 GLN CB C 32.2 0.1 1 15 . 4 GLN CG C 33.2 0.1 1 16 . 4 GLN CD C 180.2 0.1 1 17 . 4 GLN C C 174.1 0.1 1 18 . 5 LYS N N 115.2 0.1 1 19 . 5 LYS CA C 53.5 0.1 1 20 . 5 LYS CB C 33.2 0.1 1 21 . 5 LYS CG C 24.5 0.1 1 22 . 5 LYS CD C 28.8 0.1 1 23 . 5 LYS CE C 40.1 0.1 1 24 . 5 LYS C C 174.9 0.1 1 25 . 6 VAL N N 117.4 0.1 1 26 . 6 VAL CA C 55.5 0.1 1 27 . 6 VAL CB C 35.6 0.1 1 28 . 6 VAL CG1 C 19.8 0.1 2 29 . 6 VAL CG2 C 22.9 0.1 2 30 . 6 VAL C C 175.6 0.1 1 31 . 7 GLU N N 123.3 0.1 1 32 . 7 GLU CA C 54.5 0.1 1 33 . 7 GLU CB C 31.3 0.1 1 34 . 7 GLU CG C 36.8 0.1 1 35 . 7 GLU CD C 182.9 0.1 1 36 . 7 GLU C C 175.4 0.1 1 37 . 8 VAL N N 122.7 0.1 1 38 . 8 VAL CA C 63.7 0.1 1 39 . 8 VAL CB C 31.1 0.1 1 40 . 8 VAL CG1 C 21.6 0.1 2 41 . 8 VAL CG2 C 22.4 0.1 2 42 . 8 VAL C C 176.8 0.1 1 43 . 9 ARG N N 126.8 0.1 1 44 . 9 ARG CA C 54.1 0.1 1 45 . 9 ARG CB C 28.0 0.1 1 46 . 10 LEU N N 116.7 0.1 1 47 . 10 LEU CA C 52.9 0.1 1 48 . 10 LEU CB C 43.0 0.1 1 49 . 10 LEU CG C 25.5 0.1 1 50 . 10 LEU CD1 C 21.0 0.1 1 51 . 10 LEU CD2 C 21.0 0.1 1 52 . 11 LYS N N 118.6 0.1 1 53 . 11 LYS CA C 58.1 0.1 1 54 . 11 LYS CB C 33.3 0.1 1 55 . 11 LYS C C 174.1 0.1 1 56 . 12 THR N N 109.5 0.1 1 57 . 12 THR CA C 59.1 0.1 1 58 . 12 THR CB C 73.4 0.1 1 59 . 12 THR CG2 C 20.7 0.1 1 60 . 12 THR C C 173.8 0.1 1 61 . 13 GLY N N 106.4 0.1 1 62 . 13 GLY CA C 44.9 0.1 1 63 . 13 GLY C C 172.1 0.1 1 64 . 14 LEU N N 120.0 0.1 1 65 . 14 LEU CA C 56.2 0.1 1 66 . 14 LEU CB C 43.4 0.1 1 67 . 14 LEU CG C 26.6 0.1 1 68 . 14 LEU CD1 C 24.8 0.1 1 69 . 14 LEU CD2 C 24.8 0.1 1 70 . 14 LEU C C 177.2 0.1 1 71 . 15 GLN N N 118.4 0.1 1 72 . 15 GLN CA C 55.3 0.1 1 73 . 15 GLN CB C 33.8 0.1 1 74 . 15 GLN CG C 32.1 0.1 1 75 . 15 GLN CD C 180.1 0.1 1 76 . 15 GLN C C 174.1 0.1 1 77 . 16 ALA N N 118.2 0.1 1 78 . 16 ALA CA C 54.9 0.1 1 79 . 16 ALA CB C 16.9 0.1 1 80 . 16 ALA C C 179.1 0.1 1 81 . 17 ARG N N 121.4 0.1 1 82 . 17 ARG CA C 61.4 0.1 1 83 . 17 ARG CB C 27.2 0.1 1 84 . 17 ARG C C 177.5 0.1 1 85 . 18 PRO N N 133.6 0.1 1 86 . 18 PRO CA C 65.1 0.1 1 87 . 18 PRO CB C 31.6 0.1 1 88 . 18 PRO CG C 28.2 0.1 1 89 . 18 PRO CD C 49.2 0.1 1 90 . 18 PRO C C 180.0 0.1 1 91 . 19 ALA N N 119.8 0.1 1 92 . 19 ALA CA C 55.5 0.1 1 93 . 19 ALA CB C 18.3 0.1 1 94 . 20 ALA N N 121.5 0.1 1 95 . 20 ALA CA C 54.8 0.1 1 96 . 20 ALA CB C 18.3 0.1 1 97 . 21 LEU N N 119.6 0.1 1 98 . 21 LEU CA C 56.9 0.1 1 99 . 21 LEU CB C 40.6 0.1 1 100 . 21 LEU CG C 26.9 0.1 1 101 . 21 LEU CD1 C 20.8 0.1 1 102 . 21 LEU CD2 C 20.8 0.1 1 103 . 21 LEU C C 178.6 0.1 1 104 . 22 PHE N N 122.6 0.1 1 105 . 22 PHE CA C 61.1 0.1 1 106 . 22 PHE CB C 38.4 0.1 1 107 . 22 PHE CG C 138.9 0.1 1 108 . 22 PHE CD1 C 131.6 0.1 1 109 . 22 PHE CD2 C 131.6 0.1 1 110 . 22 PHE C C 177.6 0.1 1 111 . 23 VAL N N 120.2 0.1 1 112 . 23 VAL CA C 66.6 0.1 1 113 . 23 VAL CB C 31.5 0.1 1 114 . 23 VAL CG1 C 22.0 0.1 2 115 . 23 VAL CG2 C 23.3 0.1 2 116 . 23 VAL C C 177.0 0.1 1 117 . 24 GLN N N 118.2 0.1 1 118 . 24 GLN CA C 58.7 0.1 1 119 . 24 GLN CB C 29.0 0.1 1 120 . 24 GLN CG C 34.2 0.1 1 121 . 24 GLN CD C 180.1 0.1 1 122 . 25 GLU N N 117.9 0.1 1 123 . 25 GLU CA C 57.8 0.1 1 124 . 25 GLU CB C 28.6 0.1 1 125 . 25 GLU CG C 34.4 0.1 1 126 . 25 GLU CD C 182.7 0.1 1 127 . 26 ALA N N 118.2 0.1 1 128 . 26 ALA CA C 54.6 0.1 1 129 . 26 ALA CB C 17.8 0.1 1 130 . 27 ASN N N 113.9 0.1 1 131 . 27 ASN CA C 53.8 0.1 1 132 . 27 ASN CB C 38.4 0.1 1 133 . 27 ASN CG C 176.1 0.1 1 134 . 27 ASN ND2 N 110.3 0.1 1 135 . 28 ARG N N 117.9 0.1 1 136 . 28 ARG CA C 57.2 0.1 1 137 . 28 ARG CB C 28.9 0.1 1 138 . 28 ARG CG C 27.4 0.1 1 139 . 28 ARG CD C 42.8 0.1 1 140 . 29 PHE N N 118.5 0.1 1 141 . 29 PHE CA C 56.8 0.1 1 142 . 29 PHE CB C 39.6 0.1 1 143 . 29 PHE CG C 140.5 0.1 1 144 . 29 PHE CD1 C 131.5 0.1 1 145 . 29 PHE CD2 C 131.5 0.1 1 146 . 29 PHE C C 175.8 0.1 1 147 . 30 THR N N 119.8 0.1 1 148 . 30 THR CA C 63.8 0.1 1 149 . 30 THR CB C 68.7 0.1 1 150 . 30 THR CG2 C 21.9 0.1 1 151 . 30 THR C C 177.7 0.1 1 152 . 31 SER N N 121.6 0.1 1 153 . 31 SER CA C 61.6 0.1 1 154 . 31 SER CB C 64.9 0.1 1 155 . 31 SER C C 173.7 0.1 1 156 . 32 ASP N N 121.5 0.1 1 157 . 32 ASP CA C 54.2 0.1 1 158 . 32 ASP CB C 42.2 0.1 1 159 . 32 ASP CG C 180.2 0.1 1 160 . 32 ASP C C 175.5 0.1 1 161 . 33 VAL N N 120.8 0.1 1 162 . 33 VAL CA C 59.6 0.1 1 163 . 33 VAL CB C 35.2 0.1 1 164 . 33 VAL CG1 C 22.2 0.1 2 165 . 33 VAL CG2 C 20.0 0.1 2 166 . 33 VAL C C 173.9 0.1 1 167 . 34 PHE N N 122.0 0.1 1 168 . 34 PHE CA C 55.0 0.1 1 169 . 34 PHE CB C 43.1 0.1 1 170 . 34 PHE CG C 138.7 0.1 1 171 . 34 PHE CD1 C 131.6 0.1 1 172 . 34 PHE CD2 C 131.6 0.1 1 173 . 34 PHE C C 174.9 0.1 1 174 . 35 LEU N N 117.7 0.1 1 175 . 35 LEU CA C 53.2 0.1 1 176 . 35 LEU CB C 46.3 0.1 1 177 . 35 LEU CG C 27.6 0.1 1 178 . 35 LEU CD1 C 24.2 0.1 2 179 . 35 LEU CD2 C 25.4 0.1 2 180 . 35 LEU C C 175.6 0.1 1 181 . 36 GLU N N 122.9 0.1 1 182 . 36 GLU CA C 53.8 0.1 1 183 . 36 GLU CB C 34.3 0.1 1 184 . 36 GLU CG C 36.3 0.1 1 185 . 36 GLU CD C 182.0 0.1 1 186 . 36 GLU C C 174.7 0.1 1 187 . 37 LYS N N 123.8 0.1 1 188 . 37 LYS CA C 55.8 0.1 1 189 . 37 LYS CB C 36.1 0.1 1 190 . 37 LYS CG C 25.0 0.1 1 191 . 37 LYS CD C 28.9 0.1 1 192 . 37 LYS CE C 42.1 0.1 1 193 . 38 ASP N N 129.9 0.1 1 194 . 38 ASP CA C 55.4 0.1 1 195 . 38 ASP CB C 39.6 0.1 1 196 . 38 ASP CG C 180.9 0.1 1 197 . 38 ASP C C 175.5 0.1 1 198 . 39 GLY N N 105.0 0.1 1 199 . 39 GLY CA C 45.0 0.1 1 200 . 39 GLY C C 173.8 0.1 1 201 . 40 LYS N N 122.6 0.1 1 202 . 40 LYS CA C 54.0 0.1 1 203 . 40 LYS CB C 33.3 0.1 1 204 . 40 LYS CG C 25.0 0.1 1 205 . 40 LYS CD C 28.9 0.1 1 206 . 40 LYS CE C 42.1 0.1 1 207 . 41 LYS N N 126.0 0.1 1 208 . 41 LYS CA C 54.1 0.1 1 209 . 41 LYS CB C 35.4 0.1 1 210 . 41 LYS CG C 25.2 0.1 1 211 . 41 LYS CD C 28.9 0.1 1 212 . 41 LYS CE C 40.7 0.1 1 213 . 42 VAL N N 118.8 0.1 1 214 . 42 VAL CA C 57.9 0.1 1 215 . 42 VAL CB C 35.0 0.1 1 216 . 42 VAL CG1 C 18.4 0.1 2 217 . 42 VAL CG2 C 21.6 0.1 2 218 . 43 ASN N N 121.4 0.1 1 219 . 43 ASN CA C 53.0 0.1 1 220 . 43 ASN CB C 38.1 0.1 1 221 . 43 ASN CG C 176.7 0.1 1 222 . 43 ASN C C 174.9 0.1 1 223 . 43 ASN ND2 N 111.0 0.1 1 224 . 44 ALA N N 128.4 0.1 1 225 . 44 ALA CA C 51.8 0.1 1 226 . 44 ALA CB C 18.9 0.1 1 227 . 44 ALA C C 173.8 0.1 1 228 . 45 LYS N N 110.2 0.1 1 229 . 45 LYS CA C 55.6 0.1 1 230 . 45 LYS CB C 31.5 0.1 1 231 . 45 LYS CG C 27.4 0.1 1 232 . 45 LYS CD C 29.5 0.1 1 233 . 45 LYS CE C 42.8 0.1 1 234 . 45 LYS C C 175.5 0.1 1 235 . 46 SER N N 114.9 0.1 1 236 . 46 SER CA C 54.2 0.1 1 237 . 46 SER CB C 64.6 0.1 1 238 . 46 SER C C 174.4 0.1 1 239 . 47 ILE N N 131.0 0.1 1 240 . 47 ILE CA C 66.4 0.1 1 241 . 47 ILE CB C 37.7 0.1 1 242 . 47 ILE CG1 C 30.4 0.1 1 243 . 47 ILE CG2 C 17.8 0.1 1 244 . 47 ILE CD1 C 14.1 0.1 1 245 . 47 ILE C C 178.0 0.1 1 246 . 48 MET N N 117.7 0.1 1 247 . 48 MET CA C 58.3 0.1 1 248 . 48 MET CB C 32.3 0.1 1 249 . 48 MET CG C 32.2 0.1 1 250 . 48 MET CE C 15.9 0.1 1 251 . 49 GLY N N 107.9 0.1 1 252 . 49 GLY CA C 46.9 0.1 1 253 . 49 GLY C C 176.9 0.1 1 254 . 50 LEU N N 124.8 0.1 1 255 . 50 LEU CA C 58.2 0.1 1 256 . 50 LEU CB C 43.3 0.1 1 257 . 50 LEU C C 179.7 0.1 1 258 . 51 MET N N 114.3 0.1 1 259 . 51 MET CA C 59.1 0.1 1 260 . 51 MET CB C 33.6 0.1 1 261 . 51 MET CG C 33.5 0.1 1 262 . 51 MET CE C 15.8 0.1 1 263 . 52 SER N N 112.3 0.1 1 264 . 52 SER CA C 59.8 0.1 1 265 . 52 SER CB C 63.8 0.1 1 266 . 52 SER C C 175.2 0.1 1 267 . 53 LEU N N 122.0 0.1 1 268 . 53 LEU CA C 54.2 0.1 1 269 . 53 LEU CB C 42.8 0.1 1 270 . 53 LEU CG C 26.1 0.1 1 271 . 53 LEU CD1 C 22.4 0.1 1 272 . 53 LEU CD2 C 22.4 0.1 1 273 . 54 ALA N N 131.2 0.1 1 274 . 54 ALA CA C 51.8 0.1 1 275 . 54 ALA CB C 17.1 0.1 1 276 . 54 ALA C C 176.8 0.1 1 277 . 55 VAL N N 121.2 0.1 1 278 . 55 VAL CA C 60.2 0.1 1 279 . 55 VAL CB C 37.1 0.1 1 280 . 55 VAL CG1 C 20.0 0.1 2 281 . 55 VAL CG2 C 22.9 0.1 2 282 . 55 VAL C C 180.9 0.1 1 283 . 56 SER N N 118.0 0.1 1 284 . 56 SER CA C 56.7 0.1 1 285 . 56 SER CB C 65.9 0.1 1 286 . 56 SER C C 173.5 0.1 1 287 . 57 THR N N 122.4 0.1 1 288 . 57 THR CA C 65.8 0.1 1 289 . 57 THR CB C 68.4 0.1 1 290 . 57 THR CG2 C 22.4 0.1 1 291 . 57 THR C C 175.8 0.1 1 292 . 58 GLY N N 117.1 0.1 1 293 . 58 GLY CA C 44.4 0.1 1 294 . 58 GLY C C 173.9 0.1 1 295 . 59 THR N N 117.2 0.1 1 296 . 59 THR CA C 63.0 0.1 1 297 . 59 THR CB C 68.8 0.1 1 298 . 59 THR CG2 C 22.4 0.1 1 299 . 59 THR C C 172.4 0.1 1 300 . 60 GLU N N 121.8 0.1 1 301 . 60 GLU CA C 54.3 0.1 1 302 . 60 GLU CB C 32.7 0.1 1 303 . 60 GLU CG C 36.3 0.1 1 304 . 60 GLU CD C 183.2 0.1 1 305 . 61 VAL N N 118.9 0.1 1 306 . 61 VAL CA C 58.4 0.1 1 307 . 61 VAL CB C 34.8 0.1 1 308 . 61 VAL CG1 C 22.9 0.1 2 309 . 61 VAL CG2 C 19.8 0.1 2 310 . 62 THR N N 117.2 0.1 1 311 . 62 THR CA C 62.6 0.1 1 312 . 62 THR CB C 69.1 0.1 1 313 . 62 THR CG2 C 22.5 0.1 1 314 . 62 THR C C 172.6 0.1 1 315 . 63 LEU N N 131.0 0.1 1 316 . 63 LEU CA C 53.8 0.1 1 317 . 63 LEU CB C 45.0 0.1 1 318 . 63 LEU CG C 27.6 0.1 1 319 . 63 LEU CD1 C 24.2 0.1 1 320 . 63 LEU CD2 C 24.2 0.1 1 321 . 64 ILE N N 125.8 0.1 1 322 . 64 ILE CA C 59.8 0.1 1 323 . 64 ILE CB C 42.6 0.1 1 324 . 64 ILE CG1 C 28.0 0.1 1 325 . 64 ILE CG2 C 17.8 0.1 1 326 . 64 ILE CD1 C 15.0 0.1 1 327 . 64 ILE C C 173.9 0.1 1 328 . 65 ALA N N 126.4 0.1 1 329 . 65 ALA CA C 50.0 0.1 1 330 . 65 ALA CB C 23.0 0.1 1 331 . 65 ALA C C 175.9 0.1 1 332 . 66 GLN N N 120.1 0.1 1 333 . 66 GLN CA C 53.7 0.1 1 334 . 66 GLN CB C 32.1 0.1 1 335 . 66 GLN CG C 33.4 0.1 1 336 . 66 GLN CD C 179.1 0.1 1 337 . 66 GLN C C 174.2 0.1 1 338 . 67 GLY N N 119.5 0.1 1 339 . 67 GLY CA C 45.2 0.1 1 340 . 67 GLY C C 174.9 0.1 1 341 . 68 GLU N N 118.7 0.1 1 342 . 68 GLU CA C 59.2 0.1 1 343 . 68 GLU CB C 29.9 0.1 1 344 . 68 GLU CG C 36.2 0.1 1 345 . 68 GLU CD C 183.3 0.1 1 346 . 69 ASP N N 114.8 0.1 1 347 . 69 ASP CA C 51.1 0.1 1 348 . 69 ASP CB C 39.4 0.1 1 349 . 69 ASP CG C 179.6 0.1 1 350 . 69 ASP C C 176.2 0.1 1 351 . 70 GLU N N 116.3 0.1 1 352 . 70 GLU CA C 57.4 0.1 1 353 . 70 GLU CB C 27.2 0.1 1 354 . 70 GLU CG C 36.9 0.1 1 355 . 70 GLU CD C 184.5 0.1 1 356 . 70 GLU C C 175.1 0.1 1 357 . 71 GLN N N 121.4 0.1 1 358 . 71 GLN CA C 59.5 0.1 1 359 . 71 GLN CB C 27.7 0.1 1 360 . 71 GLN CG C 33.8 0.1 1 361 . 71 GLN CD C 179.7 0.1 1 362 . 72 GLU N N 117.1 0.1 1 363 . 72 GLU CA C 59.3 0.1 1 364 . 72 GLU CB C 26.8 0.1 1 365 . 72 GLU CG C 34.2 0.1 1 366 . 72 GLU CD C 183.2 0.1 1 367 . 73 ALA N N 122.4 0.1 1 368 . 73 ALA CA C 53.5 0.1 1 369 . 73 ALA CB C 17.9 0.1 1 370 . 73 ALA C C 178.7 0.1 1 371 . 74 LEU N N 116.5 0.1 1 372 . 74 LEU CA C 58.0 0.1 1 373 . 74 LEU CB C 42.1 0.1 1 374 . 74 LEU CG C 26.0 0.1 1 375 . 74 LEU CD1 C 24.3 0.1 1 376 . 74 LEU CD2 C 24.3 0.1 1 377 . 75 GLU N N 116.9 0.1 1 378 . 75 GLU CA C 58.9 0.1 1 379 . 75 GLU CB C 29.4 0.1 1 380 . 75 GLU CG C 36.2 0.1 1 381 . 75 GLU CD C 182.6 0.1 1 382 . 75 GLU C C 176.2 0.1 1 383 . 76 LYS N N 118.9 0.1 1 384 . 76 LYS CA C 59.0 0.1 1 385 . 76 LYS CB C 32.2 0.1 1 386 . 76 LYS CG C 24.8 0.1 1 387 . 76 LYS CD C 28.6 0.1 1 388 . 76 LYS CE C 42.1 0.1 1 389 . 77 LEU N N 119.3 0.1 1 390 . 77 LEU CA C 57.5 0.1 1 391 . 77 LEU CB C 40.9 0.1 1 392 . 77 LEU CG C 26.8 0.1 1 393 . 77 LEU CD1 C 22.5 0.1 1 394 . 77 LEU CD2 C 22.5 0.1 1 395 . 77 LEU C C 179.1 0.1 1 396 . 78 ALA N N 121.1 0.1 1 397 . 78 ALA CA C 55.1 0.1 1 398 . 78 ALA CB C 18.2 0.1 1 399 . 78 ALA C C 181.5 0.1 1 400 . 79 ALA N N 120.1 0.1 1 401 . 79 ALA CA C 55.6 0.1 1 402 . 79 ALA CB C 18.7 0.1 1 403 . 79 ALA C C 180.9 0.1 1 404 . 80 TYR N N 116.8 0.1 1 405 . 80 TYR CA C 61.3 0.1 1 406 . 80 TYR CB C 38.0 0.1 1 407 . 80 TYR CG C 129.7 0.1 1 408 . 80 TYR CD1 C 132.8 0.1 1 409 . 80 TYR CD2 C 132.8 0.1 1 410 . 80 TYR CE1 C 118.2 0.1 1 411 . 80 TYR CE2 C 118.2 0.1 1 412 . 80 TYR CZ C 157.3 0.1 1 413 . 80 TYR C C 176.8 0.1 1 414 . 81 VAL N N 117.8 0.1 1 415 . 81 VAL CA C 64.6 0.1 1 416 . 81 VAL CB C 32.2 0.1 1 417 . 81 VAL CG1 C 22.1 0.1 2 418 . 81 VAL CG2 C 23.5 0.1 2 419 . 81 VAL C C 174.6 0.1 1 420 . 82 GLN N N 113.4 0.1 1 421 . 82 GLN CA C 55.7 0.1 1 422 . 82 GLN CB C 29.9 0.1 1 423 . 82 GLN CG C 35.1 0.1 1 424 . 82 GLN CD C 179.0 0.1 1 425 . 83 GLU N N 114.9 0.1 1 426 . 83 GLU CA C 55.7 0.1 1 427 . 83 GLU CB C 30.0 0.1 1 428 . 83 GLU CG C 36.2 0.1 1 429 . 83 GLU CD C 181.7 0.1 1 430 . 84 GLU N N 114.9 0.1 1 431 . 84 GLU CA C 55.3 0.1 1 432 . 84 GLU CB C 29.8 0.1 1 433 . 84 GLU CG C 36.2 0.1 1 434 . 84 GLU CD C 181.7 0.1 1 435 . 85 VAL N N 93.4 0.1 1 436 . 85 VAL CA C 60.6 0.1 1 437 . 85 VAL CB C 32.7 0.1 1 438 . 85 VAL CG2 C 19.3 0.1 2 439 . 85 VAL CG1 C 21.3 0.1 2 440 . 85 VAL C C 171.1 0.1 1 stop_ save_ save_shift_set_2 _Saveframe_category assigned_chemical_shifts _Details ; Peak doubling has been observed for a limited number of signals, indicating conformational or dynamic disorder. The minor signals are reported in this list. ; loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $Ex-cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'Crh subunit 1' _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 . 10 LEU CA C 53.1 0.1 1 2 . 10 LEU CB C 42.6 0.1 1 3 . 12 THR N N 111.0 0.1 1 4 . 18 PRO C C 178.8 0.1 1 5 . 38 ASP CB C 40.0 0.1 1 6 . 38 ASP CG C 181.7 0.1 1 7 . 49 GLY N N 109.5 0.1 1 8 . 52 SER N N 113.1 0.1 1 9 . 54 ALA C C 175.7 0.1 1 10 . 67 GLY N N 117.1 0.1 1 stop_ save_