Mechanism for the selective interaction of C-terminal EH-domain proteins with specific NPF-containing partners
GPLGSDDVEW VVGKDKPTYD EIFYTLSPVN GKITGANAKK EMVKSKLPNT VLGKIWKLAD VDKDGLLDDE EFALANHLIK VKLEGHELPA DLPPHLVPPS KRRHE
Polymer type: polypeptide(L)
Total | 1H | 13C | 15N | |
---|---|---|---|---|
All | 81.9 % (1161 of 1417) | 91.8 % (683 of 744) | 69.1 % (385 of 557) | 80.2 % (93 of 116) |
Backbone | 73.5 % (513 of 698) | 89.9 % (213 of 237) | 60.2 % (212 of 352) | 80.7 % (88 of 109) |
Sidechain | 89.3 % (742 of 831) | 92.7 % (470 of 507) | 84.2 % (267 of 317) | 71.4 % (5 of 7) |
Aromatic | 86.2 % (81 of 94) | 97.9 % (46 of 47) | 75.6 % (34 of 45) | 50.0 % (1 of 2) |
Methyl | 95.3 % (122 of 128) | 96.9 % (62 of 64) | 93.8 % (60 of 64) |
1. EH domain of EHD1
GPLGSDDVEW VVGKDKPTYD EIFYTLSPVN GKITGANAKK EMVKSKLPNT VLGKIWKLAD VDKDGLLDDE EFALANHLIK VKLEGHELPA DLPPHLVPPS KRRHE2. MICAL L1 like peptide
LESKPYNPFE EEEEDSolvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7.0, Details 20mM Tris, 100Kcl, 2mM Cacl2
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EH domain of EHD1 | [U-99% 13C; U-99% 15N] | 0.6 mM | |
2 | MICAL L1 like peptide | natural abundance | 2 mM | |
3 | Tris | natural abundance | 20 mM | |
4 | Kcl | natural abundance | 100 mM | |
5 | Cac12 | natural abundance | 2 mM | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Varian INOVA - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7.0, Details 20mM Tris, 100Kcl, 2mM Cacl2
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EH domain of EHD1 | [U-99% 13C; U-99% 15N] | 0.6 mM | |
2 | MICAL L1 like peptide | natural abundance | 2 mM | |
3 | Tris | natural abundance | 20 mM | |
4 | Kcl | natural abundance | 100 mM | |
5 | Cac12 | natural abundance | 2 mM | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Varian INOVA - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7.0, Details 20mM Tris, 100Kcl, 2mM Cacl2
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EH domain of EHD1 | [U-99% 13C; U-99% 15N] | 0.6 mM | |
2 | MICAL L1 like peptide | natural abundance | 2 mM | |
3 | Tris | natural abundance | 20 mM | |
4 | Kcl | natural abundance | 100 mM | |
5 | Cac12 | natural abundance | 2 mM | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Varian INOVA - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7.0, Details 20mM Tris, 100Kcl, 2mM Cacl2
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EH domain of EHD1 | [U-99% 13C; U-99% 15N] | 0.6 mM | |
2 | MICAL L1 like peptide | natural abundance | 2 mM | |
3 | Tris | natural abundance | 20 mM | |
4 | Kcl | natural abundance | 100 mM | |
5 | Cac12 | natural abundance | 2 mM | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Varian INOVA - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7.0, Details 20mM Tris, 100Kcl, 2mM Cacl2
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EH domain of EHD1 | [U-99% 13C; U-99% 15N] | 0.6 mM | |
2 | MICAL L1 like peptide | natural abundance | 2 mM | |
3 | Tris | natural abundance | 20 mM | |
4 | Kcl | natural abundance | 100 mM | |
5 | Cac12 | natural abundance | 2 mM | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Varian INOVA - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7.0, Details 20mM Tris, 100Kcl, 2mM Cacl2
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EH domain of EHD1 | [U-99% 13C; U-99% 15N] | 0.6 mM | |
2 | MICAL L1 like peptide | natural abundance | 2 mM | |
3 | Tris | natural abundance | 20 mM | |
4 | Kcl | natural abundance | 100 mM | |
5 | Cac12 | natural abundance | 2 mM | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Varian INOVA - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7.0, Details 20mM Tris, 100Kcl, 2mM Cacl2
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EH domain of EHD1 | [U-99% 13C; U-99% 15N] | 0.6 mM | |
2 | MICAL L1 like peptide | natural abundance | 2 mM | |
3 | Tris | natural abundance | 20 mM | |
4 | Kcl | natural abundance | 100 mM | |
5 | Cac12 | natural abundance | 2 mM | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Varian INOVA - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7.0, Details 20mM Tris, 100Kcl, 2mM Cacl2
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EH domain of EHD1 | [U-99% 13C; U-99% 15N] | 0.6 mM | |
2 | MICAL L1 like peptide | natural abundance | 2 mM | |
3 | Tris | natural abundance | 20 mM | |
4 | Kcl | natural abundance | 100 mM | |
5 | Cac12 | natural abundance | 2 mM | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Varian INOVA - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7.0, Details 20mM Tris, 100Kcl, 2mM Cacl2
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EH domain of EHD1 | [U-99% 13C; U-99% 15N] | 0.6 mM | |
2 | MICAL L1 like peptide | natural abundance | 2 mM | |
3 | Tris | natural abundance | 20 mM | |
4 | Kcl | natural abundance | 100 mM | |
5 | Cac12 | natural abundance | 2 mM | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Properties
Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints | combined_16671_2ksp.nef |
Input source #2: Coordindates | 2ksp.cif |
Diamagnetism of the molecular assembly | False (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…) |
---|---|---|---|---|
1:66:LEU:O | 3:1:CA:CA | unknown | unknown | n/a |
1:71:GLU:OE2 | 3:1:CA:CA | unknown | unknown | n/a |
Non-standard residues
Chain_ID | Seq_ID | Comp_ID | Chem_comp_name | Experimental evidences |
---|---|---|---|---|
C | 1 | CA | CALCIUM ION | None |
Sequence alignments
----40--------50--------60--------70--------80--------90-------100-------110-------120-------130---- GPLGSDDVEWVVGKDKPTYDEIFYTLSPVNGKITGANAKKEMVKSKLPNTVLGKIWKLADVDKDGLLDDEEFALANHLIKVKLEGHELPADLPPHLVPPS |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| GPLGSDDVEWVVGKDKPTYDEIFYTLSPVNGKITGANAKKEMVKSKLPNTVLGKIWKLADVDKDGLLDDEEFALANHLIKVKLEGHELPADLPPHLVPPS --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 ----- KRRHE ||||| KRRHE -----
-----150------- LESKPYNPFEEEEED ||||||||||||||| LESKPYNPFEEEEED --------10-----
Chain assignments
Entity_assembly_ID (NMR) | Auth_asym_ID (model) | Length | Unmapped | Conflict | Sequence coverage (%) |
---|---|---|---|---|---|
A | A | 105 | 0 | 0 | 100.0 |
B | B | 15 | 0 | 0 | 100.0 |
Content subtype: combined_16671_2ksp.nef
Assigned chemical shifts
----40--------50--------60--------70--------80--------90-------100-------110-------120-------130---- GPLGSDDVEWVVGKDKPTYDEIFYTLSPVNGKITGANAKKEMVKSKLPNTVLGKIWKLADVDKDGLLDDEEFALANHLIKVKLEGHELPADLPPHLVPPS ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| .....DDVEWVVGKDKPTYDEIFYTLSPVNGKITGANAKKEMVKSKLPNTVLGKIWKLADVDKDGLLDDEEFALANHLIKVKLEGHELPADLPPHLVPPS ----- KRRHE ||||| KRRHE
-----150------- LESKPYNPFEEEEED | |||||||| |||| L.SKPYNPFE.EEED
Comp_index_ID | Comp_ID | Atom_ID | CS value (ppm) |
---|---|---|---|
59 | THR | HG1 | 6.04 |
68 | THR | HG1 | 5.4 |
136 | ARG | HH11 | 6.4 |
136 | ARG | HH12 | 7.16 |
136 | ARG | HH21 | 6.3 |
136 | ARG | HH22 | 7.16 |
137 | ARG | HH11 | 6.4 |
137 | ARG | HH12 | 7.16 |
137 | ARG | HH21 | 6.4 |
137 | ARG | HH22 | 7.16 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 646 | 616 | 95.4 |
13C chemical shifts | 487 | 365 | 74.9 |
15N chemical shifts | 104 | 92 | 88.5 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 209 | 197 | 94.3 |
13C chemical shifts | 210 | 99 | 47.1 |
15N chemical shifts | 96 | 87 | 90.6 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 437 | 419 | 95.9 |
13C chemical shifts | 277 | 266 | 96.0 |
15N chemical shifts | 8 | 5 | 62.5 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 63 | 61 | 96.8 |
13C chemical shifts | 63 | 61 | 96.8 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 38 | 37 | 97.4 |
13C chemical shifts | 36 | 34 | 94.4 |
15N chemical shifts | 2 | 1 | 50.0 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 98 | 71 | 72.4 |
13C chemical shifts | 70 | 0 | 0.0 |
15N chemical shifts | 14 | 0 | 0.0 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 28 | 17 | 60.7 |
13C chemical shifts | 30 | 0 | 0.0 |
15N chemical shifts | 13 | 0 | 0.0 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 70 | 54 | 77.1 |
13C chemical shifts | 40 | 0 | 0.0 |
15N chemical shifts | 1 | 0 | 0.0 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 2 | 2 | 100.0 |
13C chemical shifts | 2 | 0 | 0.0 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 9 | 9 | 100.0 |
13C chemical shifts | 9 | 0 | 0.0 |
Covalent bonds
Distance restraints
----40--------50--------60--------70--------80--------90-------100-------110-------120-------130---- GPLGSDDVEWVVGKDKPTYDEIFYTLSPVNGKITGANAKKEMVKSKLPNTVLGKIWKLADVDKDGLLDDEEFALANHLIKVKLEGHELPADLPPHLVPPS ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| .....DDVEWVVGKDKPTYDEIFYTLSPVNGKITGANAKKEMVKSKLPNTVLGKIWKLADVDKDGLLDDEEFALANHLIKVKLEGHELPADLPPHLVPPS ----- KRRHE ||||| KRRHE
-----150------- LESKPYNPFEEEEED | |||||||| |||| L.SKPYNPFE.EEED