NMR Solution Structures of butyryl-ACP (a non-polar, non pathway intermediate) from the actinorhodin polyketide synthase in Streptomyces coelicolor
Polymer type: polypeptide(L)
Total | 1H | 13C | 15N | |
---|---|---|---|---|
All | 95.9 % (874 of 911) | 94.8 % (438 of 462) | 96.7 % (352 of 364) | 98.8 % (84 of 85) |
Backbone | 96.9 % (496 of 512) | 96.6 % (171 of 177) | 96.4 % (242 of 251) | 98.8 % (83 of 84) |
Sidechain | 95.4 % (456 of 478) | 94.0 % (268 of 285) | 97.4 % (187 of 192) | 100.0 % (1 of 1) |
Aromatic | 100.0 % (36 of 36) | 100.0 % (18 of 18) | 100.0 % (18 of 18) | |
Methyl | 96.7 % (116 of 120) | 96.7 % (58 of 60) | 96.7 % (58 of 60) |
1. butyryl-ACP
MATLLTTDDL RRALVESAGE TDGTDLSGDF LDLRFEDIGY DSLALMETAA RLESRYGVSI PDDVAGRVDT PRELLDLING ALAEAASolvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | water | protons | 4.78 ppm | internal | indirect | 0.2514495 |
1H | water | protons | 4.78 ppm | internal | direct | 1.0 |
15N | water | protons | 4.78 ppm | internal | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | water | protons | 4.78 ppm | internal | indirect | 0.2514495 |
1H | water | protons | 4.78 ppm | internal | direct | 1.0 |
15N | water | protons | 4.78 ppm | internal | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | water | protons | 4.78 ppm | internal | indirect | 0.2514495 |
1H | water | protons | 4.78 ppm | internal | direct | 1.0 |
15N | water | protons | 4.78 ppm | internal | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | water | protons | 4.78 ppm | internal | indirect | 0.2514495 |
1H | water | protons | 4.78 ppm | internal | direct | 1.0 |
15N | water | protons | 4.78 ppm | internal | indirect | 0.1013291 |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Varian INOVA - 600 MHz Equipped with a triple resonance probe and XYZ PFGs.
State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 (±0.2) K, pH 5.5, Details Samples were 1mM NaN3, 5%/95% D2O/H2O in 600μL potassium phosphate buffer (20mM, pH 5.5) and 1-2mM derivatised ACP. The 4'-PP sidechain and butyryl group were not isotopically enriched
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | ACP | [U-98% 13C; U-98% 15N] | 1.0 ~ 2.0 (±0.2) mM | |
2 | potassium phosphate | natural abundance | 20 (±2.0) mM | |
3 | sodium azide | natural abundance | 1 (±0.1) mM | |
4 | H2O | natural abundance | 95 % | |
5 | D2O | natural abundance | 5 % |
Properties
Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints | combined_16199_2kg9.nef |
Input source #2: Coordindates | 2kg9.cif |
Diamagnetism of the molecular assembly | True (excluding Oxygen atoms) |
Whether the assembly has a disulfide bond | None |
Whether the assembly has a other bond | True (see coodinates for details) |
Whether the assembly contains a cyclic polymer | None |
Overall data processing status | Warning |
Disulfide bonds
NoneOther bonds (neither disulfide, covalent nor hydrogen bonds, e.g. Zinc–sulphur bond)
Ptnr_site_1 | Ptnr_site_2 | Redox_state_prediction_1 | Redox_state_prediction_2 | Distance (Ã…) |
---|---|---|---|---|
1:42:SER:OG | 2:1:PSR:P24 | unknown | unknown | n/a |
Non-standard residues
Chain_ID | Seq_ID | Comp_ID | Chem_comp_name | Experimental evidences |
---|---|---|---|---|
B | 1 | PSR | THIOBUTYRIC ACID S-{2-[3-(2-HYDROXY-3,3-DIMETHYL-4-PHOSPHONOOXY-BUTYRYLAMINO)-PROPIONYLAMINO]-ETHYL} ESTER | Assigned chemical shifts |
Sequence alignments
--------10--------20--------30--------40--------50--------60--------70--------80------ MATLLTTDDLRRALVESAGETDGTDLSGDFLDLRFEDIGYDSLALMETAARLESRYGVSIPDDVAGRVDTPRELLDLINGALAEAA |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| MATLLTTDDLRRALVESAGETDGTDLSGDFLDLRFEDIGYDSLALMETAARLESRYGVSIPDDVAGRVDTPRELLDLINGALAEAA
Chain assignments
Entity_assembly_ID (NMR) | Auth_asym_ID (model) | Length | Unmapped | Conflict | Sequence coverage (%) |
---|---|---|---|---|---|
A | A | 86 | 0 | 0 | 100.0 |
Content subtype: combined_16199_2kg9.nef
Assigned chemical shifts
--------10--------20--------30--------40--------50--------60--------70--------80------ MATLLTTDDLRRALVESAGETDGTDLSGDFLDLRFEDIGYDSLALMETAARLESRYGVSIPDDVAGRVDTPRELLDLINGALAEAA ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| .ATLLTTDDLRRALVESAGETDGTDLSGDFLDLRFEDIGYDSLALMETAARLESRYGVSIPDDVAGRVDTPRELLDLINGALAEAA
- X | X
Comp_index_ID | Comp_ID | Atom_ID | CS value (ppm) |
---|---|---|---|
11 | ARG | CZ | 159.522 |
12 | ARG | CZ | 159.629 |
34 | ARG | CZ | 159.939 |
51 | ARG | CZ | 159.659 |
55 | ARG | CZ | 159.965 |
67 | ARG | CZ | 159.707 |
72 | ARG | HH11 | 6.892 |
72 | ARG | HH12 | 6.892 |
72 | ARG | HH21 | 6.892 |
72 | ARG | HH22 | 6.892 |
72 | ARG | CZ | 159.409 |
72 | ARG | NH1 | 71.207 |
72 | ARG | NH2 | 71.207 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 462 | 453 | 98.1 |
13C chemical shifts | 364 | 354 | 97.3 |
15N chemical shifts | 92 | 90 | 97.8 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 177 | 174 | 98.3 |
13C chemical shifts | 172 | 166 | 96.5 |
15N chemical shifts | 84 | 82 | 97.6 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 285 | 279 | 97.9 |
13C chemical shifts | 192 | 188 | 97.9 |
15N chemical shifts | 8 | 8 | 100.0 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 62 | 60 | 96.8 |
13C chemical shifts | 62 | 60 | 96.8 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 18 | 18 | 100.0 |
13C chemical shifts | 18 | 18 | 100.0 |
Comp_index_ID | Comp_ID |
---|---|
1 | PSR |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|
Covalent bonds
Distance restraints
--------10--------20--------30--------40--------50--------60--------70--------80------ MATLLTTDDLRRALVESAGETDGTDLSGDFLDLRFEDIGYDSLALMETAARLESRYGVSIPDDVAGRVDTPRELLDLINGALAEAA ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| .ATLLTTDDLRRALVESAGETDGTDLSGDFLDLRFEDIGYDSLALMETAARLESRYGVSIPDDVAGRVDTPRELLDLINGALAEAA
Dihedral angle restraints
--------10--------20--------30--------40--------50--------60--------70--------80------ MATLLTTDDLRRALVESAGETDGTDLSGDFLDLRFEDIGYDSLALMETAARLESRYGVSIPDDVAGRVDTPRELLDLINGALAEAA ||||||||||||||||| ||||||||||||||||||||||||||||||||||||| |||||||||||||||| .ATLLTTDDLRRALVESA...........FLDLRFEDIGYDSLALMETAARLESRYGVSIPDDVAG..DTPRELLDLINGALAE --------10--------20--------30--------40--------50--------60--------70--------80----