data_18104 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; MARCKS MH2 Domain free and mAb 3C3 bounded ; _BMRB_accession_number 18104 _BMRB_flat_file_name bmr18104.str _Entry_type original _Submission_date 2011-11-28 _Accession_date 2011-11-28 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details ; The S25p and S25up (1.2 mM) NMR samples were prepared in 10 mM phosphate buffer saline (PBS) pH 7.2 and pH 5.0 with 10% of D2O. For the peptide-antibody interaction studies, mAb 3C3 was added directly on the NMR samples for a final concentration of 3.6 M (300:1 molar ratio peptide-antibody). NMR experiments were carried out on Varian Inova 600 MHz or on Bruker Avance DRX 600 MHz at 5 oC or 25 oC. ; loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Tinoco Luzineide W. . 2 Fraga Jully L. . 3 AnoBom Cristiane D. . 4 Zolessi Flavio . . 5 Obal Gonzalo . . 6 Toledo Andrea . . 7 Pritsch Otto . . 8 Arruti Cristina . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 91 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2014-03-14 original author . stop_ loop_ _Related_BMRB_accession_number _Relationship 18105 'S25p (phosphorylated)' 18106 'S25p complex with 3C3' stop_ _Original_release_date 2014-03-14 save_ ############################# # Citation for this entry # ############################# save_citation_1 _Saveframe_category entry_citation _Citation_full . _Citation_title 'Structural characterization of a neuroblast-specific phosphorylated region of MARCKS' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 24590112 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Tinoco Luzineide W. . 2 Fraga Jully L. . 3 AnoBom Cristiane D. . 4 Zolessi Flavio . . 5 Obal Gonzalo . . 6 Toledo Andrea . . 7 Pritsch Otto . . 8 Arruti Cristina . . stop_ _Journal_abbreviation 'Biochim. Biophys. Acta' _Journal_name_full 'Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics' _Journal_volume 1844 _Journal_issue 4 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 837 _Page_last 849 _Year 2014 _Details . loop_ _Keyword 'MH2 domain' neuroblast peptide-antibody phophorylation stop_ save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name S25up _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label S25up $MARCKS_peptides stop_ _System_molecular_weight 172000 _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _Database_query_date . _Details 'S25p (phosphorylated MARCKS peptide) interacting wiht the monoclonal antibody 3C3' save_ ######################## # Monomeric polymers # ######################## save_MARCKS_peptides _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common MARCKS_peptides _Molecular_mass 1941 _Mol_thiol_state 'not present' loop_ _Biological_function 'regions of MARCKS phosphorylated isoform specific for neuroblasts and some types of neurons' stop_ _Details P16527 ############################## # Polymer residue sequence # ############################## _Residue_count 20 _Mol_residue_sequence ; EKPGEAVAASPSKANGQENG ; loop_ _Residue_seq_code _Residue_label 1 GLU 2 LYS 3 PRO 4 GLY 5 GLU 6 ALA 7 VAL 8 ALA 9 ALA 10 SER 11 PRO 12 SER 13 LYS 14 ALA 15 ASN 16 GLY 17 GLN 18 GLU 19 ASN 20 GLY stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-11-18 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 EMBL CAJ81538 "myristoylated alanine-rich protein kinase C substrate [Xenopus (Silurana) tropicalis]" 100.00 286 100.00 100.00 1.82e-02 GB AAA48946 "C kinase substrate [Gallus gallus]" 100.00 281 100.00 100.00 1.99e-02 GB AAH75394 "marcks protein [Xenopus (Silurana) tropicalis]" 100.00 286 100.00 100.00 1.90e-02 GB EMP38065 "Myristoylated alanine-rich C-kinase substrate [Chelonia mydas]" 100.00 282 100.00 100.00 2.16e-02 GB EOB08830 "Myristoylated alanine-rich C-kinase substrate, partial [Anas platyrhynchos]" 85.00 33 100.00 100.00 5.08e-01 GB KQL91641 "myristoylated alanine-rich protein kinase C substrate [Alligator mississippiensis]" 100.00 197 100.00 100.00 1.56e-02 REF NP_001006693 "myristoylated alanine-rich C-kinase substrate [Xenopus (Silurana) tropicalis]" 100.00 286 100.00 100.00 1.82e-02 REF NP_990811 "myristoylated alanine-rich C-kinase substrate [Gallus gallus]" 100.00 281 100.00 100.00 1.99e-02 REF XP_005287641 "PREDICTED: LOW QUALITY PROTEIN: myristoylated alanine-rich C-kinase substrate [Chrysemys picta bellii]" 100.00 280 100.00 100.00 2.24e-02 REF XP_005518342 "PREDICTED: myristoylated alanine-rich C-kinase substrate [Pseudopodoces humilis]" 100.00 286 100.00 100.00 2.45e-02 REF XP_005518343 "PREDICTED: myristoylated alanine-rich C-kinase substrate [Pseudopodoces humilis]" 100.00 286 100.00 100.00 2.45e-02 SP P16527 "RecName: Full=Myristoylated alanine-rich C-kinase substrate; Short=MARCKS [Gallus gallus]" 100.00 281 100.00 100.00 1.99e-02 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $MARCKS_peptides chicken 9031 Eukaryota Metazoa Gallus gallus 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 $MARCKS_peptides 'chemical synthesis' . . . . . stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_S25up _Saveframe_category sample _Sample_type solution _Details 'MARCKS unphosphorylated peptide' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $MARCKS_peptides 1.2 mM 'natural abundance' 'sodium phosphate' 10 mM '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_NMRView _Saveframe_category software _Name NMRView _Version . loop_ _Vendor _Address _Electronic_address 'Johnson, One Moon Scientific' . . stop_ loop_ _Task 'chemical shift assignment' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DRX _Field_strength 600 _Details . save_ save_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model INOVA _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-1H_TOCSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label $sample_S25up save_ save_2D_1H-1H_NOESY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $sample_S25up save_ save_2D_1H-1H_TOCSY _Saveframe_category NMR_applied_experiment _Experiment_name 2D_1H-1H_TOCSY _BMRB_pulse_sequence_accession_number . _Details ; Water suppression was achieved using WATERGATE pulse sequence. TOCSY spectra were acquired using MLEV-17 for spin lock, using a mixing time of 70 ms and the spectra were collected with 256 data points in F1 and 2048 points in F2 with 32 transients/t1. ; save_ save_2D_1H-1H_NOESY _Saveframe_category NMR_applied_experiment _Experiment_name 2D_1H-1H_NOESY _BMRB_pulse_sequence_accession_number . _Details 'For NOESY spectra, an optimal mixing time of 300 ms was used, observed in a range of 150-500 ms.' save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 13.7 . mM pH 7.2 . pH pressure 1 . atm temperature 298 . K stop_ save_ save_sample_conditions_2 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 13.7 . mM pH 7.2 . pH pressure 1 . atm temperature 278 . K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference_1 _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 water H 1 protons ppm 4.7 internal direct . . . 1.0 stop_ save_ save_chemical_shift_reference_2 _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 water H 1 protons ppm 4.96 internal direct . . . 1.0 stop_ save_ ################################### # Assigned chemical shift lists # ################################### ################################################################### # Chemical Shift Ambiguity Index Value Definitions # # # # The values other than 1 are used for those atoms with different # # chemical shifts that cannot be assigned to stereospecific atoms # # or to specific residues or chains. # # # # Index Value Definition # # # # 1 Unique (including isolated methyl protons, # # geminal atoms, and geminal methyl # # groups with identical chemical shifts) # # (e.g. ILE HD11, HD12, HD13 protons) # # 2 Ambiguity of geminal atoms or geminal methyl # # proton groups (e.g. ASP HB2 and HB3 # # protons, LEU CD1 and CD2 carbons, or # # LEU HD11, HD12, HD13 and HD21, HD22, # # HD23 methyl protons) # # 3 Aromatic atoms on opposite sides of # # symmetrical rings (e.g. TYR HE1 and HE2 # # protons) # # 4 Intraresidue ambiguities (e.g. LYS HG and # # HD protons or TRP HZ2 and HZ3 protons) # # 5 Interresidue ambiguities (LYS 12 vs. LYS 27) # # 6 Intermolecular ambiguities (e.g. ASP 31 CA # # in monomer 1 and ASP 31 CA in monomer 2 # # of an asymmetrical homodimer, duplex # # DNA assignments, or other assignments # # that may apply to atoms in one or more # # molecule in the molecular assembly) # # 9 Ambiguous, specific ambiguity not defined # # # ################################################################### save_S25up-25 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-1H TOCSY' '2D 1H-1H NOESY' stop_ loop_ _Sample_label $sample_S25up stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name S25up _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 2 LYS HA H 4.565 . 1 2 2 2 LYS HB3 H 1.745 . 2 3 2 2 LYS HG2 H 1.376 . 2 4 2 2 LYS HD2 H 1.605 . 2 5 2 2 LYS HE2 H 2.905 . 2 6 3 3 PRO HA H 4.346 . 1 7 3 3 PRO HB2 H 2.361 . 2 8 3 3 PRO HB3 H 2.234 . 2 9 3 3 PRO HG2 H 1.929 . 2 10 3 3 PRO HG3 H 1.975 . 2 11 3 3 PRO HD2 H 3.889 . 2 12 3 3 PRO HD3 H 3.575 . 2 13 5 5 GLU H H 8.519 . 1 14 5 5 GLU HA H 4.166 . 1 15 5 5 GLU HB2 H 1.854 . 2 16 5 5 GLU HB3 H 1.941 . 2 17 5 5 GLU HG2 H 2.167 . 2 18 6 6 ALA H H 8.258 . 1 19 6 6 ALA HA H 4.208 . 1 20 6 6 ALA HB H 1.291 . 1 21 6 6 ALA HB H 1.273 . 1 22 6 6 ALA HB H 1.291 . 1 23 7 7 VAL H H 7.986 . 1 24 7 7 VAL HA H 3.971 . 1 25 7 7 VAL HB H 1.951 . 1 26 7 7 VAL HG1 H 0.838 . 2 27 7 7 VAL HG1 H 0.877 . 2 28 7 7 VAL HG2 H 0.838 . 2 29 7 7 VAL HG2 H 0.877 . 2 30 8 8 ALA H H 8.256 . 1 31 8 8 ALA HA H 4.229 . 1 32 8 8 ALA HB H 1.293 . 1 33 8 8 ALA HB H 1.266 . 1 34 8 8 ALA HB H 1.293 . 1 35 9 9 ALA H H 8.163 . 1 36 9 9 ALA HB H 1.298 . 1 37 9 9 ALA HB H 1.266 . 1 38 9 9 ALA HB H 1.298 . 1 39 10 10 SER H H 8.477 . 9 40 10 10 SER HA H 4.595 . 1 41 10 10 SER HB3 H 3.850 . 2 42 11 11 PRO HA H 4.370 . 1 43 11 11 PRO HB2 H 2.210 . 2 44 11 11 PRO HB3 H 2.231 . 2 45 11 11 PRO HG2 H 1.930 . 2 46 11 11 PRO HG3 H 1.850 . 2 47 11 11 PRO HD2 H 3.641 . 2 48 11 11 PRO HD3 H 3.729 . 2 49 12 12 SER H H 8.282 . 1 50 12 12 SER HA H 4.660 . 1 51 12 12 SER HB2 H 3.789 . 2 52 12 12 SER HB3 H 3.874 . 2 53 13 13 LYS H H 8.216 . 1 54 13 13 LYS HA H 4.235 . 1 55 13 13 LYS HB3 H 1.754 . 2 56 13 13 LYS HG2 H 1.356 . 2 57 13 13 LYS HG3 H 1.337 . 2 58 13 13 LYS HD2 H 1.584 . 2 59 13 13 LYS HD3 H 1.650 . 2 60 13 13 LYS HE2 H 2.899 . 2 61 14 14 ALA H H 8.162 . 1 62 14 14 ALA HA H 4.219 . 1 63 14 14 ALA HB H 1.310 . 1 64 14 14 ALA HB H 1.270 . 1 65 14 14 ALA HB H 1.310 . 1 66 15 15 ASN H H 8.326 . 1 67 15 15 ASN HA H 4.657 . 1 68 15 15 ASN HB2 H 2.752 . 2 69 15 15 ASN HB3 H 2.705 . 2 70 15 15 ASN HD21 H 7.342 . 2 71 15 15 ASN HD22 H 7.433 . 2 72 16 16 GLY H H 8.278 . 1 73 16 16 GLY HA2 H 3.829 . 2 74 17 17 GLN H H 8.124 . 1 75 17 17 GLN HA H 4.250 . 1 76 17 17 GLN HB2 H 1.871 . 1 77 17 17 GLN HB3 H 2.034 . 2 78 17 17 GLN HG2 H 2.241 . 1 79 17 17 GLN HG3 H 2.311 . 2 80 18 18 GLU H H 7.967 . 1 81 18 18 GLU HA H 4.187 . 1 82 18 18 GLU HB2 H 1.851 . 2 83 18 18 GLU HB3 H 1.959 . 2 84 18 18 GLU HG2 H 2.146 . 2 85 18 18 GLU HG3 H 2.214 . 2 86 19 19 ASN H H 8.473 . 1 87 19 19 ASN HA H 4.598 . 1 88 19 19 ASN HB2 H 2.776 . 2 89 19 19 ASN HB3 H 2.695 . 2 90 20 20 GLY H H 8.474 . 1 91 20 20 GLY HA2 H 4.150 . 2 stop_ save_