data_50591 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Ebola Virus Glycoprotein Interacts with Cholesterol to Enhance Membrane Fusion and Cell Entry, G660L mutant ; _BMRB_accession_number 50591 _BMRB_flat_file_name bmr50591.str _Entry_type original _Submission_date 2020-11-22 _Accession_date 2020-11-22 _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 Lee Jinwoo . . 2 Liang Binyong . . 3 Tamm Lukas K. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 2 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 80 "15N chemical shifts" 80 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2020-12-22 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 30162 'Envelope glycoprotein' 50584 'Ebola GP2 MPER/TM, wt' stop_ _Original_release_date 2020-11-23 save_ ############################# # Citation for this entry # ############################# save_citations_1 _Saveframe_category entry_citation _Citation_full . _Citation_title ; Ebola Virus Glycoprotein Interacts with Cholesterol to Enhance Membrane Fusion and Cell Entry ; _Citation_status 'in press' _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 Lee Jinwoo . . 2 Liang Binyong . . 3 Tamm Lukas K. . stop_ _Journal_abbreviation 'Nat. Struct. Mol. Biol.' _Journal_volume . _Journal_issue . _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first . _Page_last . _Year . _Details . loop_ _Keyword 'ebola virus, cholesterol, protein-cholesterol interaction, membrane fusion' stop_ save_ ################################## # Molecular system description # ################################## save_assembly_1 _Saveframe_category molecular_system _Mol_system_name 'Ebola GP2 MPER/TM, G660L mutant' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'Ebola GP2 MPER/TM, G660L mutant' $entity_1 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_entity_1 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common entity_1 _Molecular_mass 5188 _Mol_thiol_state 'not present' loop_ _Biological_function 'viral entry membrane fusion' stop_ _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 47 _Mol_residue_sequence ; GSDKTLPDQGDNDNWWTGWR QWIPAGIGVTLVVIAVIALF AIAKFVF ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 630 GLY 2 631 SER 3 632 ASP 4 633 LYS 5 634 THR 6 635 LEU 7 636 PRO 8 637 ASP 9 638 GLN 10 639 GLY 11 640 ASP 12 641 ASN 13 642 ASP 14 643 ASN 15 644 TRP 16 645 TRP 17 646 THR 18 647 GLY 19 648 TRP 20 649 ARG 21 650 GLN 22 651 TRP 23 652 ILE 24 653 PRO 25 654 ALA 26 655 GLY 27 656 ILE 28 657 GLY 29 658 VAL 30 659 THR 31 660 LEU 32 661 VAL 33 662 VAL 34 663 ILE 35 664 ALA 36 665 VAL 37 666 ILE 38 667 ALA 39 668 LEU 40 669 PHE 41 670 ALA 42 671 ILE 43 672 ALA 44 673 LYS 45 674 PHE 46 675 VAL 47 676 PHE stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . 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 UNP Q05320 GP . . . . . stop_ save_ #################### # Natural source # #################### save_natural_source_1 _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Strain $entity_1 'Zaire ebolavirus' 186538 Viruses . Ebolavirus ZEBOV '(strain Mayinga-76)' stop_ save_ ######################### # Experimental source # ######################### save_experimental_source_1 _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Vector_type _Vector_name $entity_1 'recombinant technology' . Escherichia coli BL21(DE3) plasmid pET24a 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 $entity_1 0.5 mM '[U-99% 13C; U-99% 15N]' 'sodium phosphate' 25 mM 'natural abundance' 'sodium chloride' 100 mM 'natural abundance' DHPC 188 mM 'natural abundance' DMPC 94 mM 'natural abundance' stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $entity_1 0.5 mM '[U-99% 13C; U-99% 15N]' Cholesterol 15 % 'natural abundance' 'sodium phosphate' 25 mM 'natural abundance' 'sodium chloride' 100 mM 'natural abundance' DHPC 188 mM 'natural abundance' DMPC 94 mM 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_software_1 _Saveframe_category software _Name NMRPipe _Version . loop_ _Task processing stop_ _Details . save_ save_software_2 _Saveframe_category software _Name SPARKY _Version . loop_ _Task 'data analysis' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model 'AVANCE III' _Field_strength 800 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_1 save_ save_2D_1H-15N_HSQC_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _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 'ionic strength' 0.1 . M pH 5.5 . pH pressure 1 . atm temperature 318 . K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chem_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 DSS H 1 'methyl protons' ppm 0 external direct . . . 1 DSS N 15 'methyl protons' ppm 0 external indirect . . . . 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_assigned_chemical_shifts_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Software_label $software_2 stop_ loop_ _Experiment_label '2D 1H-15N HSQC' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chem_shift_reference_1 _Mol_system_component_name 'Ebola GP2 MPER/TM, G660L mutant' _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 1 632 3 ASP H H 8.315 . . 2 632 3 ASP N N 122.676 . . 3 633 4 LYS H H 8.092 . . 4 633 4 LYS N N 121.759 . . 5 634 5 THR H H 8.027 . . 6 634 5 THR N N 115.784 . . 7 635 6 LEU H H 7.940 . . 8 635 6 LEU N N 125.870 . . 9 637 8 ASP H H 8.153 . . 10 637 8 ASP N N 120.277 . . 11 638 9 GLN H H 8.109 . . 12 638 9 GLN N N 120.850 . . 13 639 10 GLY H H 8.265 . . 14 639 10 GLY N N 110.072 . . 15 640 11 ASP H H 8.062 . . 16 640 11 ASP N N 120.721 . . 17 641 12 ASN H H 8.216 . . 18 641 12 ASN N N 119.121 . . 19 642 13 ASP H H 8.111 . . 20 642 13 ASP N N 120.557 . . 21 643 14 ASN H H 7.947 . . 22 643 14 ASN N N 118.768 . . 23 644 15 TRP H H 8.080 . . 24 644 15 TRP N N 122.090 . . 25 645 16 TRP H H 7.165 . . 26 645 16 TRP N N 118.120 . . 27 646 17 THR H H 7.404 . . 28 646 17 THR N N 114.044 . . 29 647 18 GLY H H 8.066 . . 30 647 18 GLY N N 110.624 . . 31 648 19 TRP H H 7.640 . . 32 648 19 TRP N N 120.903 . . 33 649 20 ARG H H 7.308 . . 34 649 20 ARG N N 119.641 . . 35 650 21 GLN H H 7.705 . . 36 650 21 GLN N N 116.056 . . 37 651 22 TRP H H 7.643 . . 38 651 22 TRP N N 118.054 . . 39 655 26 GLY H H 8.588 . . 40 655 26 GLY N N 105.820 . . 41 657 28 GLY H H 8.390 . . 42 657 28 GLY N N 108.291 . . 43 658 29 VAL H H 8.452 . . 44 658 29 VAL N N 119.461 . . 45 659 30 THR H H 7.625 . . 46 659 30 THR N N 116.823 . . 47 660 31 LEU H H 8.150 . . 48 660 31 LEU N N 119.589 . . 49 661 32 VAL H H 7.699 . . 50 661 32 VAL N N 118.151 . . 51 662 33 VAL H H 8.099 . . 52 662 33 VAL N N 119.070 . . 53 663 34 ILE H H 8.192 . . 54 663 34 ILE N N 117.987 . . 55 664 35 ALA H H 8.114 . . 56 664 35 ALA N N 121.822 . . 57 665 36 VAL H H 8.195 . . 58 665 36 VAL N N 116.664 . . 59 666 37 ILE H H 8.119 . . 60 666 37 ILE N N 119.589 . . 61 667 38 ALA H H 8.572 . . 62 667 38 ALA N N 121.279 . . 63 668 39 LEU H H 8.228 . . 64 668 39 LEU N N 117.489 . . 65 669 40 PHE H H 8.223 . . 66 669 40 PHE N N 118.481 . . 67 670 41 ALA H H 8.429 . . 68 670 41 ALA N N 120.789 . . 69 671 42 ILE H H 8.272 . . 70 671 42 ILE N N 116.446 . . 71 672 43 ALA H H 8.202 . . 72 672 43 ALA N N 121.705 . . 73 673 44 LYS H H 7.662 . . 74 673 44 LYS N N 114.430 . . 75 674 45 PHE H H 7.849 . . 76 674 45 PHE N N 115.962 . . 77 675 46 VAL H H 7.658 . . 78 675 46 VAL N N 115.760 . . 79 676 47 PHE H H 7.329 . . 80 676 47 PHE N N 124.273 . . stop_ save_ save_assigned_chemical_shifts_2 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Software_label $software_2 stop_ loop_ _Experiment_label '2D 1H-15N HSQC' stop_ loop_ _Sample_label $sample_2 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chem_shift_reference_1 _Mol_system_component_name 'Ebola GP2 MPER/TM, G660L mutant' _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 1 632 3 ASP H H 8.314 . . 2 632 3 ASP N N 122.692 . . 3 633 4 LYS H H 8.086 . . 4 633 4 LYS N N 121.753 . . 5 634 5 THR H H 8.023 . . 6 634 5 THR N N 115.812 . . 7 635 6 LEU H H 7.945 . . 8 635 6 LEU N N 125.933 . . 9 637 8 ASP H H 8.150 . . 10 637 8 ASP N N 120.301 . . 11 638 9 GLN H H 8.109 . . 12 638 9 GLN N N 120.850 . . 13 639 10 GLY H H 8.264 . . 14 639 10 GLY N N 110.084 . . 15 640 11 ASP H H 8.062 . . 16 640 11 ASP N N 120.738 . . 17 641 12 ASN H H 8.214 . . 18 641 12 ASN N N 119.143 . . 19 642 13 ASP H H 8.111 . . 20 642 13 ASP N N 120.557 . . 21 643 14 ASN H H 7.945 . . 22 643 14 ASN N N 118.775 . . 23 644 15 TRP H H 8.084 . . 24 644 15 TRP N N 122.076 . . 25 645 16 TRP H H 7.161 . . 26 645 16 TRP N N 118.132 . . 27 646 17 THR H H 7.398 . . 28 646 17 THR N N 114.018 . . 29 647 18 GLY H H 8.069 . . 30 647 18 GLY N N 110.676 . . 31 648 19 TRP H H 7.642 . . 32 648 19 TRP N N 120.940 . . 33 649 20 ARG H H 7.297 . . 34 649 20 ARG N N 119.632 . . 35 650 21 GLN H H 7.686 . . 36 650 21 GLN N N 116.021 . . 37 651 22 TRP H H 7.636 . . 38 651 22 TRP N N 118.028 . . 39 655 26 GLY H H 8.584 . . 40 655 26 GLY N N 105.797 . . 41 657 28 GLY H H 8.396 . . 42 657 28 GLY N N 108.263 . . 43 658 29 VAL H H 8.455 . . 44 658 29 VAL N N 119.440 . . 45 659 30 THR H H 7.622 . . 46 659 30 THR N N 116.803 . . 47 660 31 LEU H H 8.150 . . 48 660 31 LEU N N 119.589 . . 49 661 32 VAL H H 7.707 . . 50 661 32 VAL N N 118.184 . . 51 662 33 VAL H H 8.105 . . 52 662 33 VAL N N 119.065 . . 53 663 34 ILE H H 8.195 . . 54 663 34 ILE N N 118.001 . . 55 664 35 ALA H H 8.114 . . 56 664 35 ALA N N 121.822 . . 57 665 36 VAL H H 8.199 . . 58 665 36 VAL N N 116.672 . . 59 666 37 ILE H H 8.126 . . 60 666 37 ILE N N 119.588 . . 61 667 38 ALA H H 8.576 . . 62 667 38 ALA N N 121.296 . . 63 668 39 LEU H H 8.229 . . 64 668 39 LEU N N 117.489 . . 65 669 40 PHE H H 8.224 . . 66 669 40 PHE N N 118.480 . . 67 670 41 ALA H H 8.424 . . 68 670 41 ALA N N 120.805 . . 69 671 42 ILE H H 8.274 . . 70 671 42 ILE N N 116.452 . . 71 672 43 ALA H H 8.199 . . 72 672 43 ALA N N 121.711 . . 73 673 44 LYS H H 7.657 . . 74 673 44 LYS N N 114.432 . . 75 674 45 PHE H H 7.847 . . 76 674 45 PHE N N 115.932 . . 77 675 46 VAL H H 7.654 . . 78 675 46 VAL N N 115.724 . . 79 676 47 PHE H H 7.320 . . 80 676 47 PHE N N 124.219 . . stop_ save_