Structure, dynamics and RNA binding of the multi-domain splicing factor TIA-1
AMANHFHVFV GDLSPEITTE DIKAAFAPFG RISDARVVKD MATGKSKGYG FVSFFNKWDA ENAIQQMGGQ WLGGRQIRTN WATRKPPAPK STYESNTKQL SYDEVVNQSS PSNCTVYCGG VTSGLTEQLM RQTFSPFGQI MEIRVFPDKG YSFVRFNSHE SAAHAIVSVN GTTIEGHVVK CYWGK
Polymer type: polypeptide(L)
Total | 1H | 13C | 15N | |
---|---|---|---|---|
All | 95.4 % (2031 of 2130) | 96.2 % (1059 of 1101) | 93.3 % (774 of 830) | 99.5 % (198 of 199) |
Backbone | 97.7 % (1069 of 1094) | 97.6 % (370 of 379) | 97.2 % (523 of 538) | 99.4 % (176 of 177) |
Sidechain | 93.9 % (1130 of 1204) | 95.4 % (689 of 722) | 91.1 % (419 of 460) | 100.0 % (22 of 22) |
Aromatic | 90.3 % (213 of 236) | 90.7 % (107 of 118) | 89.5 % (102 of 114) | 100.0 % (4 of 4) |
Methyl | 98.8 % (168 of 170) | 97.6 % (83 of 85) | 100.0 % (85 of 85) |
1. TIA-1 RRM2,3
AMANHFHVFV GDLSPEITTE DIKAAFAPFG RISDARVVKD MATGKSKGYG FVSFFNKWDA ENAIQQMGGQ WLGGRQIRTN WATRKPPAPK STYESNTKQL SYDEVVNQSS PSNCTVYCGG VTSGLTEQLM RQTFSPFGQI MEIRVFPDKG YSFVRFNSHE SAAHAIVSVN GTTIEGHVVK CYWGKSolvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
8 | TIA-1 RRM2,3 | [U-99% 15N] | 0.1-0.55 mM | |
9 | sodium phosphate | natural abundance | 10 mM | |
10 | sodium chloride | natural abundance | 50 mM | |
11 | DTT | natural abundance | 10 mM | |
12 | sodium azide | natural abundance | 0.02 % | |
13 | H2O | natural abundance | 90 % | |
14 | D2O | natural abundance | 10 % |
Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
15 | TIA-1 RRM2,3 | [U-10% 13C] | 0.1-0.5 mM | |
16 | sodium phosphate | natural abundance | 10 mM | |
17 | sodium chloride | natural abundance | 50 mM | |
18 | DTT | natural abundance | 10 mM | |
19 | sodium azide | natural abundance | 0.02 % | |
20 | H2O | natural abundance | 90 % | |
21 | D2O | natural abundance | 10 % |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | na | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | na | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | na | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | na | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | na | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | na | indirect | 0.1013291 |
Atom type | Mol. common name | Atom group | Value | Ref. method | Ref. type | Shift ratio |
---|---|---|---|---|---|---|
13C | DSS | methyl protons | 0.0 ppm | na | indirect | 0.2514495 |
1H | DSS | methyl protons | 0.0 ppm | internal | direct | 1.0 |
15N | DSS | methyl protons | 0.0 ppm | na | indirect | 0.1013291 |
Bruker Avance - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
15 | TIA-1 RRM2,3 | [U-10% 13C] | 0.1-0.5 mM | |
16 | sodium phosphate | natural abundance | 10 mM | |
17 | sodium chloride | natural abundance | 50 mM | |
18 | DTT | natural abundance | 10 mM | |
19 | sodium azide | natural abundance | 0.02 % | |
20 | H2O | natural abundance | 90 % | |
21 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Bruker Avance - 900 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Bruker Avance - 750 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
8 | TIA-1 RRM2,3 | [U-99% 15N] | 0.1-0.55 mM | |
9 | sodium phosphate | natural abundance | 10 mM | |
10 | sodium chloride | natural abundance | 50 mM | |
11 | DTT | natural abundance | 10 mM | |
12 | sodium azide | natural abundance | 0.02 % | |
13 | H2O | natural abundance | 90 % | |
14 | D2O | natural abundance | 10 % |
Bruker Avance - 800 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Bruker Avance - 600 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Bruker Avance - 800 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | TIA-1 RRM2,3 | [U-99% 13C; U-99% 15N] | 0.1-0.7 mM | |
2 | sodium phosphate | natural abundance | 10 mM | |
3 | sodium chloride | natural abundance | 50 mM | |
4 | DTT | natural abundance | 10 mM | |
5 | sodium azide | natural abundance | 0.02 % | |
6 | H2O | natural abundance | 90 % | |
7 | D2O | natural abundance | 10 % |
Properties
Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints | combined_19735_2mjn.nef |
Input source #2: Coordindates | 2mjn.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
-90------100-------110-------120-------130-------140-------150-------160-------170-------180-------1 GAMANHFHVFVGDLSPEITTEDIKAAFAPFGRISDARVVKDMATGKSKGYGFVSFFNKWDAENAIQQMGGQWLGGRQIRTNWATRKPPAPKSTYESNTKQ |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| GAMANHFHVFVGDLSPEITTEDIKAAFAPFGRISDARVVKDMATGKSKGYGFVSFFNKWDAENAIQQMGGQWLGGRQIRTNWATRKPPAPKSTYESNTKQ --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 90-------200-------210-------220-------230-------240-------250-------260-------270---- LSYDEVVNQSSPSNCTVYCGGVTSGLTEQLMRQTFSPFGQIMEIRVFPDKGYSFVRFNSHESAAHAIVSVNGTTIEGHVVKCYWGK |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| LSYDEVVNQSSPSNCTVYCGGVTSGLTEQLMRQTFSPFGQIMEIRVFPDKGYSFVRFNSHESAAHAIVSVNGTTIEGHVVKCYWGK -------110-------120-------130-------140-------150-------160-------170-------180------
Chain assignments
Entity_assembly_ID (NMR) | Auth_asym_ID (model) | Length | Unmapped | Conflict | Sequence coverage (%) |
---|---|---|---|---|---|
A | A | 186 | 0 | 0 | 100.0 |
Content subtype: combined_19735_2mjn.nef
Assigned chemical shifts
-90------100-------110-------120-------130-------140-------150-------160-------170-------180-------1 GAMANHFHVFVGDLSPEITTEDIKAAFAPFGRISDARVVKDMATGKSKGYGFVSFFNKWDAENAIQQMGGQWLGGRQIRTNWATRKPPAPKSTYESNTKQ ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| .AMANHFHVFVGDLSPEITTEDIKAAFAPFGRISDARVVKDMATGKSKGYGFVSFFNKWDAENAIQQMGGQWLGGRQIRTNWATRKPPAPKSTYESNTKQ 90-------200-------210-------220-------230-------240-------250-------260-------270---- LSYDEVVNQSSPSNCTVYCGGVTSGLTEQLMRQTFSPFGQIMEIRVFPDKGYSFVRFNSHESAAHAIVSVNGTTIEGHVVKCYWGK ||||||||||||||||||| |||||||||| |||| |||||||||||| ||||||||||||||||| ||||| ||||||||| ||| LSYDEVVNQSSPSNCTVYC.GVTSGLTEQL.RQTF.PFGQIMEIRVFP.KGYSFVRFNSHESAAHA.VSVNG.TIEGHVVKC.WGK
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 1104 | 1037 | 93.9 |
13C chemical shifts | 832 | 739 | 88.8 |
15N chemical shifts | 208 | 191 | 91.8 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 382 | 363 | 95.0 |
13C chemical shifts | 372 | 341 | 91.7 |
15N chemical shifts | 178 | 169 | 94.9 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 722 | 674 | 93.4 |
13C chemical shifts | 460 | 398 | 86.5 |
15N chemical shifts | 30 | 22 | 73.3 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 90 | 86 | 95.6 |
13C chemical shifts | 90 | 86 | 95.6 |
Atom group | # of target shifts | # of assigned shifts | Completeness (%) |
---|---|---|---|
1H chemical shifts | 118 | 103 | 87.3 |
13C chemical shifts | 114 | 98 | 86.0 |
15N chemical shifts | 4 | 4 | 100.0 |
Distance restraints
-90------100-------110-------120-------130-------140-------150-------160-------170-------180-------1 GAMANHFHVFVGDLSPEITTEDIKAAFAPFGRISDARVVKDMATGKSKGYGFVSFFNKWDAENAIQQMGGQWLGGRQIRTNWATRKPPAPKSTYESNTKQ ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| ||||||||||||| ...ANHFHVFVGDLSPEITTEDIKAAFAPFGRISDARVVKDMATGKSKGYGFVSFFNKWDAENAIQQMGGQWLGGRQIRTNWATRK.PAPKSTYESNTKQ 90-------200-------210-------220-------230-------240-------250-------260-------270---- LSYDEVVNQSSPSNCTVYCGGVTSGLTEQLMRQTFSPFGQIMEIRVFPDKGYSFVRFNSHESAAHAIVSVNGTTIEGHVVKCYWGK |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| LSYDEVVNQSSPSNCTVYCGGVTSGLTEQLMRQTFSPFGQIMEIRVFPDKGYSFVRFNSHESAAHAIVSVNGTTIEGHVVKCYWGK
-90------100-------110-------120-------130-------140-------150-------160-------170-------180-------1 GAMANHFHVFVGDLSPEITTEDIKAAFAPFGRISDARVVKDMATGKSKGYGFVSFFNKWDAENAIQQMGGQWLGGRQIRTNWATRKPPAPKSTYESNTKQ | |||| || ||||||||||| || || | | | | ||||||||||||||||||||| || || | ......F.VFVG.LS..ITTEDIKAAFA..GR.SD.R.V.D...G..KGYGFVSFFNKWDAENAIQQM.GQ......IR.N................... -90------100-------110-------120-------130-------140-------150-------160-------170-------180-------1 90-------200-------210-------220-------230-------240-------250-------260-------270---- LSYDEVVNQSSPSNCTVYCGGVTSGLTEQLMRQTFSPFGQIMEIRVFPDKGYSFVRFNSHESAAHAIVSVNGTTIEGHVVKCYWGK ||| |||| | |||||||| |||||||||| | || | | ||||||| ||||||||||| | | | || | | .SYD.VVNQ.S..NCTVYCGG.....TEQLMRQTFS.F...ME.R.F...GYSFVRF..HESAAHAIVSV..T.I..H.VK.Y.G 90-------200-------210-------220-------230-------240-------250-------260-------270---
Dihedral angle restraints
-90------100-------110-------120-------130-------140-------150-------160-------170-------180-------1 GAMANHFHVFVGDLSPEITTEDIKAAFAPFGRISDARVVKDMATGKSKGYGFVSFFNKWDAENAIQQMGGQWLGGRQIRTNWATRKPPAPKSTYESNTKQ ||||||||| |||||||||||||||||||||||||||||||||| ||||||||||||||||||||| ||| |||||||| |||||| ...ANHFHVFVG.LSPEITTEDIKAAFAPFGRISDARVVKDMATGKS.GYGFVSFFNKWDAENAIQQMG..WLG.RQIRTNWA.RKPPAP.......... -90------100-------110-------120-------130-------140-------150-------160-------170-------180-------1 90-------200-------210-------220-------230-------240-------250-------260-------270---- LSYDEVVNQSSPSNCTVYCGGVTSGLTEQLMRQTFSPFGQIMEIRVFPDKGYSFVRFNSHESAAHAIVSVNGTTIEGHVVKCYWGK ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| LSYDEVVNQSSPSNCTVYCGGVTSGLTEQLMRQTFSPFGQIMEIRVFPDKGYSFVRFNSHESAAHAIVSVNGTTIEGHVVKCYWG 90-------200-------210-------220-------230-------240-------250-------260-------270---