data_26519 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Studying Invisible Excited Protein States in Slow Exchange with a Major State Conformation ; _BMRB_accession_number 26519 _BMRB_flat_file_name bmr26519.str _Entry_type original _Submission_date 2015-02-24 _Accession_date 2015-02-24 _Entry_origination BMRB _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 Vallurupalli Pramodh . . 2 Bouvignies Guillaume . . 3 Kay Lewis E. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count T1_relaxation 1 T2_relaxation 2 stop_ loop_ _Data_type _Data_type_count "T1 relaxation values" 56 "T2 relaxation values" 112 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2015-03-09 original author . stop_ loop_ _Related_BMRB_accession_number _Relationship 26520 'A39G FF domain' stop_ _Original_release_date 2015-03-09 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'Studying Invisible Excited Protein States in Slow Exchange with a Major State Conformation' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 22554188 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Vallurupalli Pramodh . . 2 Bouvignies Guillaume . . 3 Kay Lewis E. . stop_ _Journal_abbreviation 'J. Am. Chem. Soc.' _Journal_volume 134 _Journal_issue 19 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 8148 _Page_last 8161 _Year 2012 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'Abp1p-Ark1p system' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label Abp1p $Abp1p Ark1p $Ark1p stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_Abp1p _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common Abp1p _Molecular_mass . _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 59 _Mol_residue_sequence ; APWATAEYDYDAAEDNELTF VENDKIINIEFVDDDWWLGE LEKDGSKGLFPSNYVSLGN ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 1 ALA 2 2 PRO 3 3 TRP 4 4 ALA 5 5 THR 6 6 ALA 7 7 GLU 8 8 TYR 9 9 ASP 10 10 TYR 11 11 ASP 12 12 ALA 13 13 ALA 14 14 GLU 15 15 ASP 16 16 ASN 17 17 GLU 18 18 LEU 19 19 THR 20 20 PHE 21 21 VAL 22 22 GLU 23 23 ASN 24 24 ASP 25 25 LYS 26 26 ILE 27 27 ILE 28 28 ASN 29 29 ILE 30 30 GLU 31 31 PHE 32 32 VAL 33 33 ASP 34 34 ASP 35 35 ASP 36 36 TRP 37 37 TRP 38 38 LEU 39 39 GLY 40 40 GLU 41 41 LEU 42 42 GLU 43 43 LYS 44 44 ASP 45 45 GLY 46 46 SER 47 47 LYS 48 48 GLY 49 49 LEU 50 50 PHE 51 51 PRO 52 52 SER 53 53 ASN 54 54 TYR 55 55 VAL 56 56 SER 57 57 LEU 58 58 GLY 59 59 ASN stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-10-07 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 16843 Abp1p 100.00 62 100.00 100.00 5.72e-33 BMRB 18054 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18055 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18056 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18057 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18058 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18059 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18060 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18061 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18062 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18063 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18064 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18065 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18066 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18067 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18068 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18069 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18070 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18071 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18072 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18073 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18074 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18075 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18076 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18077 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 18078 AbpSH3 100.00 59 100.00 100.00 8.58e-33 BMRB 25354 Abp1p_SH3 100.00 62 100.00 100.00 5.72e-33 PDB 1JO8 "Structural Analysis Of The Yeast Actin Binding Protein Abp1 Sh3 Domain" 98.31 58 100.00 100.00 3.51e-32 PDB 2K3B "Seeing The Invisible: Structures Of Excited Protein States By Relaxation Dispersion Nmr" 100.00 62 100.00 100.00 5.72e-33 PDB 2RPN "A Crucial Role For High Intrinsic Specificity In The Function Of Yeast Sh3 Domains" 100.00 59 100.00 100.00 8.58e-33 DBJ GAA21988 "K7_Abp1p [Saccharomyces cerevisiae Kyokai no. 7]" 98.31 592 100.00 100.00 6.83e-32 EMBL CAA36075 "unnamed protein product [Saccharomyces cerevisiae]" 98.31 592 100.00 100.00 7.41e-32 EMBL CAA42253 "actin-binding protein [Saccharomyces cerevisiae]" 98.31 592 100.00 100.00 7.80e-32 EMBL CAY78291 "Abp1p [Saccharomyces cerevisiae EC1118]" 98.31 592 100.00 100.00 6.63e-32 GB AHN96147 "ABP1 [synthetic construct]" 98.31 592 100.00 100.00 7.80e-32 GB AHV79349 "ABP1 [synthetic construct]" 98.31 592 100.00 100.00 7.80e-32 GB AHY79755 "Abp1p [Saccharomyces cerevisiae YJM993]" 98.31 592 100.00 100.00 6.83e-32 GB AJP37506 "Abp1p [Saccharomyces cerevisiae YJM1078]" 98.31 592 100.00 100.00 6.83e-32 GB AJQ31991 "Abp1p [Saccharomyces cerevisiae YJM1356]" 98.31 592 100.00 100.00 7.26e-32 PRF 1603360A "actin binding protein" 98.31 592 100.00 100.00 7.57e-32 REF NP_010012 "Abp1p [Saccharomyces cerevisiae S288c]" 98.31 592 100.00 100.00 7.80e-32 SP P15891 "RecName: Full=Actin-binding protein" 98.31 592 100.00 100.00 7.80e-32 TPG DAA07557 "TPA: Abp1p [Saccharomyces cerevisiae S288c]" 98.31 592 100.00 100.00 7.80e-32 stop_ save_ save_Ark1p _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common Ark1p _Molecular_mass . _Mol_thiol_state 'not present' _Details . _Residue_count 17 _Mol_residue_sequence KKTKPTPPPKPSHLKPK loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 1 LYS 2 2 LYS 3 3 THR 4 4 LYS 5 5 PRO 6 6 THR 7 7 PRO 8 8 PRO 9 9 PRO 10 10 LYS 11 11 PRO 12 12 SER 13 13 HIS 14 14 LEU 15 15 LYS 16 16 PRO 17 17 LYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $Abp1p 'bakers yeast' 4932 Eukaryota Fungi Saccharomyces cerevisiae $Ark1p 'bakers yeast' 4932 Eukaryota Fungi Saccharomyces cerevisiae 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 _Details $Abp1p 'recombinant technology' . Escherichia coli . BL21(DE3) 'pET32b plasmid' $Ark1p 'recombinant technology' . Escherichia coli . BL21(DE3) 'pET32b plasmid' stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details 'The concentration of added peptide was such that the sample was 97.5% free, 2.5% peptide-bound' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $Abp1p 1.5 mM '[U-100% 2H; U-100% 15N]' $Ark1p 1.46 mM 'natural abundance' 'sodium phosphate' 50 mM 'natural abundance' NaCl 100 mM 'natural abundance' EDTA 1 mM 'natural abundance' NaN3 1 mM 'natural abundance' H20 90 % 'natural abundance' D2O 10 % 'natural abundance' stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $Abp1p 1.5 mM '[U-100% 2H; U-100% 15N]' $Ark1p 1.5 mM 'natural abundance' 'sodium phosphate' 50 mM 'natural abundance' NaCl 100 mM 'natural abundance' EDTA 1 mM 'natural abundance' NaN3 1 mM 'natural abundance' H20 90 % 'natural abundance' D2O 10 % 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_NMRPipe _Saveframe_category software _Name NMRPipe _Version . loop_ _Vendor _Address _Electronic_address 'Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax' . . stop_ loop_ _Task processing stop_ _Details . save_ save_SPARKY _Saveframe_category software _Name SPARKY _Version . loop_ _Vendor _Address _Electronic_address Goddard . . stop_ loop_ _Task Visualization stop_ _Details . save_ save_CATIA _Saveframe_category software _Name CATIA _Version . loop_ _Vendor _Address _Electronic_address 'Bloch McConnell' . . stop_ loop_ _Task 'data analysis' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model Inova _Field_strength 500 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-15N_HSQC_1_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC 1' _Sample_label $sample_1 save_ save_2D_1H-15N_HSQC_2_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC 2' _Sample_label $sample_2 save_ save_2D_1H-15N_HMQC_1_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HMQC 1' _Sample_label $sample_1 save_ save_2D_1H-15N_HMQC_2_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HMQC 2' _Sample_label $sample_2 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units temperature 274 . K pH 7.0 . pH pressure 1 . atm stop_ save_ save_heteronuclear_T1_list_1 _Saveframe_category T1_relaxation _Details ; Ground State Values of obtained from fits of all CEST profiles recorded on the Abp1p-Ark1p system, 1C, 11.7T to a global two-site exchange processes. Values of (kex,pE) were fixed to (130 s-1, 2.38%) and profiles for all of the residues fit simultaneously. The following six residues showed clear evidence for a second state (well resolved dips for both ground and excited states in profiles): 15 31 32 33 34 37 ; loop_ _Sample_label $sample_1 $sample_2 stop_ _Sample_conditions_label $sample_conditions_1 _Spectrometer_frequency_1H 500 _T1_coherence_type Sz _T1_value_units s-1 _Mol_system_component_name Abp1p _Text_data_format . _Text_data . loop_ _T1_ID _Residue_seq_code _Residue_label _Atom_name _T1_value _T1_value_error 1 1 ALA N 1.60 0.01 2 3 TRP N 1.87 0.01 3 4 ALA N 2.02 0.01 4 5 THR N 1.88 0.01 5 6 ALA N 1.97 0.01 6 7 GLU N 1.98 0.02 7 8 TYR N 1.92 0.01 8 9 ASP N 1.66 0.01 9 10 TYR N 1.96 0.01 10 11 ASP N 1.75 0.01 11 12 ALA N 1.81 0.01 12 13 ALA N 1.81 0.01 13 14 GLU N 1.64 0.01 14 15 ASP N 1.67 0.01 15 16 ASN N 1.79 0.01 16 17 GLU N 1.86 0.01 17 18 LEU N 2.05 0.01 18 19 THR N 1.67 0.01 19 20 PHE N 1.98 0.01 20 21 VAL N 1.86 0.01 21 22 GLU N 1.78 0.01 22 23 ASN N 1.91 0.01 23 24 ASP N 1.89 0.01 24 25 LYS N 1.77 0.01 25 27 ILE N 1.99 0.01 26 28 ASN N 1.81 0.01 27 29 ILE N 1.98 0.03 28 30 GLU N 1.87 0.01 29 31 PHE N 1.64 0.01 30 32 VAL N 1.70 0.01 31 33 ASP N 1.97 0.01 32 34 ASP N 1.70 0.01 33 35 ASP N 1.84 0.01 34 36 TRP N 1.83 0.01 35 37 TRP N 1.96 0.01 36 38 LEU N 1.90 0.01 37 39 GLY N 1.93 0.01 38 40 GLU N 1.48 0.01 39 41 LEU N 2.00 0.01 40 42 GLU N 1.96 0.01 41 43 LYS N 1.89 0.01 42 44 ASP N 1.96 0.01 43 45 GLY N 1.92 0.01 44 46 SER N 1.91 0.01 45 47 LYS N 1.70 0.01 46 48 GLY N 1.91 0.01 47 49 LEU N 1.84 0.01 48 50 PHE N 2.04 0.01 49 52 SER N 1.88 0.01 50 53 ASN N 1.85 0.01 51 54 TYR N 1.99 0.01 52 55 VAL N 1.95 0.01 53 56 SER N 1.87 0.01 54 57 LEU N 1.76 0.01 55 58 GLY N 1.91 0.01 56 59 ASN N 1.71 0.01 stop_ save_ save_heteronuclear_T2_list_G_State _Saveframe_category T2_relaxation _Details ; Ground State Values of obtained from fits of all CEST profiles recorded on the Abp1p-Ark1p system, 1C, 11.7T to a global two-site exchange processes. Values of (kex,pE) were fixed to (130 s-1, 2.38%) and profiles for all of the residues fit simultaneously. The following six residues showed clear evidence for a second state (well resolved dips for both ground and excited states in profiles): 15 31 32 33 34 37 ; loop_ _Sample_label $sample_1 $sample_2 stop_ _Sample_conditions_label $sample_conditions_1 _Spectrometer_frequency_1H 500 _T2_coherence_type S(+,-) _T2_value_units s-1 _Mol_system_component_name Abp1p _Text_data_format . _Text_data . loop_ _T2_ID _Residue_seq_code _Residue_label _Atom_name _T2_value _T2_value_error _Rex_value _Rex_error 1 1 ALA N 5.33 0.25 . . 2 3 TRP N 8.71 0.36 . . 3 4 ALA N 8.78 0.39 . . 4 5 THR N 9.63 0.30 . . 5 6 ALA N 7.09 0.45 . . 6 7 GLU N 9.28 1.39 . . 7 8 TYR N 8.87 0.25 . . 8 9 ASP N 8.48 0.23 . . 9 10 TYR N 9.57 0.24 . . 10 11 ASP N 8.52 0.21 . . 11 12 ALA N 8.46 0.32 . . 12 13 ALA N 8.72 0.25 . . 13 14 GLU N 8.05 0.22 . . 14 15 ASP N 8.27 0.10 . . 15 16 ASN N 8.18 0.19 . . 16 17 GLU N 9.64 0.14 . . 17 18 LEU N 9.49 0.34 . . 18 19 THR N 8.16 0.19 . . 19 20 PHE N 9.19 0.65 . . 20 21 VAL N 9.45 0.60 . . 21 22 GLU N 7.92 0.39 . . 22 23 ASN N 8.36 0.35 . . 23 24 ASP N 9.65 0.38 . . 24 25 LYS N 8.53 0.37 . . 25 27 ILE N 8.39 0.77 . . 26 28 ASN N 8.46 0.56 . . 27 29 ILE N 9.12 2.46 . . 28 30 GLU N 9.35 0.23 . . 29 31 PHE N 9.73 0.13 . . 30 32 VAL N 7.71 0.13 . . 31 33 ASP N 8.12 0.18 . . 32 34 ASP N 8.07 0.11 . . 33 35 ASP N 8.36 0.16 . . 34 36 TRP N 8.47 0.23 . . 35 37 TRP N 9.91 0.28 . . 36 38 LEU N 9.29 0.51 . . 37 39 GLY N 9.60 0.34 . . 38 40 GLU N 2.94 0.14 . . 39 41 LEU N 9.51 0.33 . . 40 42 GLU N 8.55 0.25 . . 41 43 LYS N 8.43 0.71 . . 42 44 ASP N 8.49 0.29 . . 43 45 GLY N 8.98 0.46 . . 44 46 SER N 8.36 0.44 . . 45 47 LYS N 7.20 0.37 . . 46 48 GLY N 8.93 0.68 . . 47 49 LEU N 9.55 0.22 . . 48 50 PHE N 9.78 0.69 . . 49 52 SER N 12.14 0.23 . . 50 53 ASN N 12.74 0.33 . . 51 54 TYR N 9.45 0.64 . . 52 55 VAL N 9.97 0.32 . . 53 56 SER N 9.46 0.42 . . 54 57 LEU N 9.26 0.38 . . 55 58 GLY N 8.89 0.46 . . 56 59 ASN N 6.60 0.37 . . stop_ save_ save_heteronuclear_T2_list_E_State _Saveframe_category T2_relaxation _Details ; Excited State Values of obtained from fits of all CEST profiles recorded on the Abp1p-Ark1p system, 1C, 11.7T to a global two-site exchange processes. Values of (kex,pE) were fixed to (130 s-1, 2.38%) and profiles for all of the residues fit simultaneously. The following six residues showed clear evidence for a second state (well resolved dips for both ground and excited states in profiles): 15 31 32 33 34 37 ; loop_ _Sample_label $sample_1 $sample_2 stop_ _Sample_conditions_label $sample_conditions_1 _Spectrometer_frequency_1H 500 _T2_coherence_type S(+,-) _T2_value_units s-1 _Mol_system_component_name Abp1p _Text_data_format . _Text_data . loop_ _T2_ID _Residue_seq_code _Residue_label _Atom_name _T2_value _T2_value_error _Rex_value _Rex_error 1 1 ALA N 0.7 10.2 . . 2 3 TRP N 4.8 14.5 . . 3 4 ALA N 2.6 15.5 . . 4 5 THR N 16.4 11.8 . . 5 6 ALA N 94.7 28.5 . . 6 7 GLU N 12.3 60.4 . . 7 8 TYR N 5.7 8.5 . . 8 9 ASP N 0.1 8.8 . . 9 10 TYR N 19.4 9.5 . . 10 11 ASP N 3.8 8.7 . . 11 12 ALA N 2.4 13.4 . . 12 13 ALA N 10.9 9.8 . . 13 14 GLU N 3.2 8.1 . . 14 15 ASP N 13.4 3.6 . . 15 16 ASN N 18.6 7.7 . . 16 17 GLU N 7.1 4.5 . . 17 18 LEU N 1.3 12.2 . . 18 19 THR N 4.8 7.6 . . 19 20 PHE N 17.1 28.3 . . 20 21 VAL N 2.9 24.9 . . 21 22 GLU N 58.4 21.5 . . 22 23 ASN N 15.2 15.7 . . 23 24 ASP N 7.4 16.3 . . 24 25 LYS N 2.6 14.8 . . 25 27 ILE N 39.7 38.3 . . 26 28 ASN N 47.8 28.9 . . 27 29 ILE N 22.7 110 . . 28 30 GLU N 3.0 7.7 . . 29 31 PHE N 6.3 4.2 . . 30 32 VAL N 19.9 5.2 . . 31 33 ASP N 13.1 6.9 . . 32 34 ASP N 27.7 4.4 . . 33 35 ASP N 22.2 6.0 . . 34 36 TRP N 12.8 8.0 . . 35 37 TRP N 4.0 8.8 . . 36 38 LEU N 2.5 19.4 . . 37 39 GLY N 18.1 13.8 . . 38 40 GLU N 0.8 5.5 . . 39 41 LEU N 2.4 13.3 . . 40 42 GLU N 0.2 10.1 . . 41 43 LYS N 18.4 33.0 . . 42 44 ASP N 1.5 11.2 . . 43 45 GLY N 0.6 18.4 . . 44 46 SER N 25.1 20.2 . . 45 47 LYS N 66.9 20.6 . . 46 48 GLY N 23.8 31.6 . . 47 49 LEU N 15.9 8.5 . . 48 50 PHE N 1.2 27.9 . . 49 52 SER N 44.7 9.1 . . 50 53 ASN N 0.4 10.1 . . 51 54 TYR N 5.9 26.8 . . 52 55 VAL N 176.8 26.0 . . 53 56 SER N 35.4 20.0 . . 54 57 LEU N 39.7 19.0 . . 55 58 GLY N 2.2 18.0 . . 56 59 ASN N 0.5 14.9 . . stop_ save_