The third member of the eIF4E family represses gene expression via a novel mode of recognition of the methyl-7 guanosine cap moiety
MALPPAAAPP GANEPLDKAL SALPPEPGGV PLHSPWTFWL DRSLPGATAA ECASNLKKIY TVQTVQIFWS VYNNIPPVTS LPLRCSYHLM RGERRPLWEE ESNAKGGVWK MKVPKDSTST VWKELLLATI GEQFTDCAAA DDEIIGVSVS VRDREDVVQV WNVNASLVGE ATVLEKIHQL LPHIAFKAVF YKPHEEHHAF EGGRGKH
Polymer type: polypeptide(L)
Total | 1H | 13C | 15N | |
---|---|---|---|---|
All | 81.8 % (1957 of 2393) | 88.2 % (1092 of 1238) | 69.5 % (658 of 947) | 99.5 % (207 of 208) |
Backbone | 82.6 % (996 of 1206) | 97.3 % (398 of 409) | 67.4 % (410 of 608) | 99.5 % (188 of 189) |
Sidechain | 82.8 % (1143 of 1381) | 83.7 % (694 of 829) | 80.7 % (430 of 533) | 100.0 % (19 of 19) |
Aromatic | 63.0 % (131 of 208) | 72.1 % (75 of 104) | 50.5 % (49 of 97) | 100.0 % (7 of 7) |
Methyl | 97.5 % (236 of 242) | 99.2 % (120 of 121) | 95.9 % (116 of 121) |
1. EUKARYOTIC TRANSLATION INITIATION FACTOR 4E TYPE
MALPPAAAPP GANEPLDKAL SALPPEPGGV PLHSPWTFWL DRSLPGATAA ECASNLKKIY TVQTVQIFWS VYNNIPPVTS LPLRCSYHLM RGERRPLWEE ESNAKGGVWK MKVPKDSTST VWKELLLATI GEQFTDCAAA DDEIIGVSVS VRDREDVVQV WNVNASLVGE ATVLEKIHQL LPHIAFKAVF YKPHEEHHAF EGGRGKHSolvent system 50mM Phosphate, 100mM NaCl, 100uM TCEP, NaN3, 7% D2O, Pressure 1.000 atm, Temperature 293.000 K, pH 7.300, Details eIF4E3 0.2mM
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EUKARYOTIC TRANSLATION INITIATION FACTOR 4E TYPE | [U-13C; U-15N] | 0.2 mM | |
2 | Phosphate | natural abundance | 50 mM | |
3 | NaCl | natural abundance | 100 mM | |
4 | TCEP | natural abundance | 100 uM | |
5 | D2O | [U-100% 2H] | 7 % |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | external | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | external | indirect | 1.0 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | external | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | external | indirect | 1.0 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | external | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | external | indirect | 1.0 |
Varian UnityInova - 800 MHz
State isotropic, Solvent system 50mM Phosphate, 100mM NaCl, 100uM TCEP, NaN3, 7% D2O, Pressure 1.000 atm, Temperature 293.000 K, pH 7.300, Details eIF4E3 0.2mM
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | EUKARYOTIC TRANSLATION INITIATION FACTOR 4E TYPE | [U-13C; U-15N] | 0.2 mM | |
2 | Phosphate | natural abundance | 50 mM | |
3 | NaCl | natural abundance | 100 mM | |
4 | TCEP | natural abundance | 100 uM | |
5 | D2O | [U-100% 2H] | 7 % |
Properties
Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints, RDC restraints | combined_18663_4b6v.nef |
Input source #2: Coordindates | 4b6v.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 | None |
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)
NoneNon-standard residues
NoneSequence alignments
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 MALPPAAAPPGANEPLDKALSALPPEPGGVPLHSPWTFWLDRSLPGATAAECASNLKKIYTVQTVQIFWSVYNNIPPVTSLPLRCSYHLMRGERRPLWEE |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| MALPPAAAPPGANEPLDKALSALPPEPGGVPLHSPWTFWLDRSLPGATAAECASNLKKIYTVQTVQIFWSVYNNIPPVTSLPLRCSYHLMRGERRPLWEE -------110-------120-------130-------140-------150-------160-------170-------180-------190-------200 ESNAKGGVWKMKVPKDSTSTVWKELLLATIGEQFTDCAAADDEIIGVSVSVRDREDVVQVWNVNASLVGEATVLEKIHQLLPHIAFKAVFYKPHEEHHAF |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| ESNAKGGVWKMKVPKDSTSTVWKELLLATIGEQFTDCAAADDEIIGVSVSVRDREDVVQVWNVNASLVGEATVLEKIHQLLPHIAFKAVFYKPHEEHHAF ------- EGGRGKH ||||||| EGGRGKH
Chain assignments
Entity_assembly_ID (NMR) | Auth_asym_ID (model) | Length | Unmapped | Conflict | Sequence coverage (%) |
---|---|---|---|---|---|
A | A | 207 | 0 | 0 | 100.0 |
Content subtype: combined_18663_4b6v.nef
Assigned chemical shifts
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 MALPPAAAPPGANEPLDKALSALPPEPGGVPLHSPWTFWLDRSLPGATAAECASNLKKIYTVQTVQIFWSVYNNIPPVTSLPLRCSYHLMRGERRPLWEE |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| MALPPAAAPPGANEPLDKALSALPPEPGGVPLHSPWTFWLDRSLPGATAAECASNLKKIYTVQTVQIFWSVYNNIPPVTSLPLRCSYHLMRGERRPLWEE -------110-------120-------130-------140-------150-------160-------170-------180-------190-------200 ESNAKGGVWKMKVPKDSTSTVWKELLLATIGEQFTDCAAADDEIIGVSVSVRDREDVVQVWNVNASLVGEATVLEKIHQLLPHIAFKAVFYKPHEEHHAF ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| || ESNAKGGVWKMKVPKDSTSTVWKELLLATIGEQFTDCAAADDEIIGVSVSVRDREDVVQVWNVNASLVGEATVLEKIHQLLPHIAFKAVFYKPHEEH.AF ------- EGGRGKH ||||||| EGGRGKH
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 1238 | 1083 | 87.5 |
13C chemical shifts | 947 | 620 | 65.5 |
15N chemical shifts | 216 | 207 | 95.8 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 409 | 399 | 97.6 |
13C chemical shifts | 414 | 197 | 47.6 |
15N chemical shifts | 189 | 188 | 99.5 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 829 | 684 | 82.5 |
13C chemical shifts | 533 | 423 | 79.4 |
15N chemical shifts | 27 | 19 | 70.4 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 124 | 122 | 98.4 |
13C chemical shifts | 124 | 117 | 94.4 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 104 | 66 | 63.5 |
13C chemical shifts | 97 | 44 | 45.4 |
15N chemical shifts | 7 | 7 | 100.0 |
Distance restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 MALPPAAAPPGANEPLDKALSALPPEPGGVPLHSPWTFWLDRSLPGATAAECASNLKKIYTVQTVQIFWSVYNNIPPVTSLPLRCSYHLMRGERRPLWEE ||| ||||||||||||| ||||||||| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| MAL........ANEPLDKALSALP.EPGGVPLHS.WTFWLDRSLPGATAAECASNLKKIYTVQTVQIFWSVYNNIPPVTSLPLRCSYHLMRGERRPLWEE -------110-------120-------130-------140-------150-------160-------170-------180-------190-------200 ESNAKGGVWKMKVPKDSTSTVWKELLLATIGEQFTDCAAADDEIIGVSVSVRDREDVVQVWNVNASLVGEATVLEKIHQLLPHIAFKAVFYKPHEEHHAF |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| |||| || ESNAKGGVWKMKVPKDSTSTVWKELLLATIGEQFTDCAAADDEIIGVSVSVRDREDVVQVWNVNASLVGEATVLEKIHQLLPHIAFKAVFYK.HEEH.AF ------- EGGRGKH || || EG...KH
Dihedral angle restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 MALPPAAAPPGANEPLDKALSALPPEPGGVPLHSPWTFWLDRSLPGATAAECASNLKKIYTVQTVQIFWSVYNNIPPVTSLPLRCSYHLMRGERRPLWEE |||||||||||||||||||||||||| |||||||||||||||||||||||||||||||||||| ||||||||||||||||| .................KALSALPPEPGGVPLHSPWTFWLDRS...ATAAECASNLKKIYTVQTVQIFWSVYNNIPPVTSLP.RCSYHLMRGERRPLWEE --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 -------110-------120-------130-------140-------150-------160-------170-------180-------190-------200 ESNAKGGVWKMKVPKDSTSTVWKELLLATIGEQFTDCAAADDEIIGVSVSVRDREDVVQVWNVNASLVGEATVLEKIHQLLPHIAFKAVFYKPHEEHHAF |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| ESNAKGGVWKMKVPKDSTSTVWKELLLATIGEQFTDCAAADDEIIGVSVSVRDREDVVQVWNVNASLVGEATVLEKIHQLLPHIAFKAVFYKPHEEHH -------110-------120-------130-------140-------150-------160-------170-------180-------190-------- ------- EGGRGKH
RDC restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 MALPPAAAPPGANEPLDKALSALPPEPGGVPLHSPWTFWLDRSLPGATAAECASNLKKIYTVQTVQIFWSVYNNIPPVTSLPLRCSYHLMRGERRPLWEE ||| || ||| | |||||||| ||||| | ||| ||| ||| | | |||| ||| || ...............................LHS.WT.WLD..L.GATAAECA..LKKIY.V.TVQ......NNI...TSL..R.S.HLMR.ERR.LW.. --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 -------110-------120-------130-------140-------150-------160-------170-------180-------190-------200 ESNAKGGVWKMKVPKDSTSTVWKELLLATIGEQFTDCAAADDEIIGVSVSVRDREDVVQVWNVNASLVGEATVLEKIHQLLPHIAFKAVFYKPHEEHHAF ||| || |||| |||||| | ||||| || | | |||||||||||||| |||| || ||||||||| ||||| || | || | ||| | ESN.KG.VWKM...KDSTST.W.ELLLA.IG.Q.T.CAAADDEIIGVSVS.RDRE.VV...NVNASLVGE.TVLEK.HQ.L.HI..K..FYK..E -------110-------120-------130-------140-------150-------160-------170-------180-------190----- ------- EGGRGKH