Structure of an engineered splicing intein mutant based on Mycobacterium tuberculosis RecA
CLAEGTRIFD PVTGTTHRIE DVVGGRKPIH VVAAAKDGTL HARPVVSWFD QGTRDVIGLR IAGGAILWAT PDHKVLTEYG WRAAGELRKG DRVAVRDVET GELRYSVIRE VLPTRRARTF DLEVEELHTL VAEGVVVHN
Polymer type: polypeptide(L)
Total | 1H | 13C | 15N | |
---|---|---|---|---|
All | 75.1 % (1170 of 1557) | 89.3 % (712 of 797) | 51.9 % (322 of 621) | 97.8 % (136 of 139) |
Backbone | 81.6 % (672 of 824) | 98.3 % (282 of 287) | 64.3 % (259 of 403) | 97.8 % (131 of 134) |
Sidechain | 70.6 % (606 of 858) | 84.3 % (430 of 510) | 49.9 % (171 of 343) | 100.0 % (5 of 5) |
Aromatic | 40.6 % (43 of 106) | 75.5 % (40 of 53) | 0.0 % (0 of 50) | 100.0 % (3 of 3) |
Methyl | 70.7 % (140 of 198) | 94.9 % (94 of 99) | 46.5 % (46 of 99) |
1. RecA
CLAEGTRIFD PVTGTTHRIE DVVGGRKPIH VVAAAKDGTL HARPVVSWFD QGTRDVIGLR IAGGAILWAT PDHKVLTEYG WRAAGELRKG DRVAVRDVET GELRYSVIRE VLPTRRARTF DLEVEELHTL VAEGVVVHNSolvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | sodium phosphate | natural abundance | 50 mM | |
2 | sodium chloride | natural abundance | 100 mM | |
3 | sodium azide | natural abundance | 1 mM | |
4 | entity | [U-98% 13C; U-98% 15N] | 0.15 ~ 0.2 mM | |
5 | H2O | natural abundance | 90 % | |
6 | 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 |
Bruker ARX - 800 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | sodium phosphate | natural abundance | 50 mM | |
2 | sodium chloride | natural abundance | 100 mM | |
3 | sodium azide | natural abundance | 1 mM | |
4 | entity | [U-98% 13C; U-98% 15N] | 0.15 ~ 0.2 mM | |
5 | H2O | natural abundance | 90 % | |
6 | D2O | natural abundance | 10 % |
Bruker ARX - 800 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | sodium phosphate | natural abundance | 50 mM | |
2 | sodium chloride | natural abundance | 100 mM | |
3 | sodium azide | natural abundance | 1 mM | |
4 | entity | [U-98% 13C; U-98% 15N] | 0.15 ~ 0.2 mM | |
5 | H2O | natural abundance | 90 % | |
6 | D2O | natural abundance | 10 % |
Bruker ARX - 800 MHz
State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 7
# | Name | Isotope labeling | Type | Concentration |
---|---|---|---|---|
1 | sodium phosphate | natural abundance | 50 mM | |
2 | sodium chloride | natural abundance | 100 mM | |
3 | sodium azide | natural abundance | 1 mM | |
4 | entity | [U-98% 13C; U-98% 15N] | 0.15 ~ 0.2 mM | |
5 | H2O | natural abundance | 90 % | |
6 | D2O | natural abundance | 10 % |
Properties
Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints, RDC restraints | combined_17414_2l8l.nef |
Input source #2: Coordindates | 2l8l.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 | Error |
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 CLAEGTRIFDPVTGTTHRIEDVVDGRKPIHVVAAAKDGTLHARPVVSWFDQGTRDVIGLRIAGGAILWATPDHKVLTEYGWRAAGELRKGDRVAVRDVET |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| CLAEGTRIFDPVTGTTHRIEDVVDGRKPIHVVAAAKDGTLHARPVVSWFDQGTRDVIGLRIAGGAILWATPDHKVLTEYGWRAAGELRKGDRVAVRDVET -------110-------120-------130--------- GELRYSVIREVLPTRRARTFDLEVEELHTLVAEGVVVHN ||||||||||||||||||||||||||||||||||||||| GELRYSVIREVLPTRRARTFDLEVEELHTLVAEGVVVHN
Chain assignments
Entity_assembly_ID (NMR) | Auth_asym_ID (model) | Length | Unmapped | Conflict | Sequence coverage (%) |
---|---|---|---|---|---|
A | A | 139 | 0 | 0 | 100.0 |
Content subtype: combined_17414_2l8l.nef
Assigned chemical shifts
Distance restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 CLAEGTRIFDPVTGTTHRIEDVVDGRKPIHVVAAAKDGTLHARPVVSWFDQGTRDVIGLRIAGGAILWATPDHKVLTEYGWRAAGELRKGDRVAVRDVET |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| CLAEGTRIFDPVTGTTHRIEDVVDGRKPIHVVAAAKDGTLHARPVVSWFDQGTRDVIGLRIAGGAILWATPDHKVLTEYGWRAAGELRKGDRVAVRDVET -------110-------120-------130--------- GELRYSVIREVLPTRRARTFDLEVEELHTLVAEGVVVHN ||||||||||||||||||||||||||||||||||||||| GELRYSVIREVLPTRRARTFDLEVEELHTLVAEGVVVHN
Dihedral angle restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 CLAEGTRIFDPVTGTTHRIEDVVDGRKPIHVVAAAKDGTLHARPVVSWFDQGTRDVIGLRIAGGAILWATPDHKVLTEYGWRAAGELRKGDRVAVRDVET ||||||||||||| ||||||||||| ||||||||||||||||| |||| ||||||||| ||||||||||||||| |||||| |||||||||||||| CLAEGTRIFDPVT.TTHRIEDVVDG....HVVAAAKDGTLHARPVV.WFDQ.TRDVIGLRI..GAILWATPDHKVLTE.GWRAAG.LRKGDRVAVRDVET --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 -------110-------120-------130--------- GELRYSVIREVLPTRRARTFDLEVEELHTLVAEGVVVHN ||||||||||||||||||||||||||| |||||||||| GELRYSVIREVLPTRRARTFDLEVEEL.TLVAEGVVVH -------110-------120-------130--------
RDC restraints
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 CLAEGTRIFDPVTGTTHRIEDVVDGRKPIHVVAAAKDGTLHARPVVSWFDQGTRDVIGLRIAGGAILWATPDHKVLTEYGWRAAGELRKGDRVAVRDVET | ||| || ||||||||||||||| | ||| | ||||||| || || || | ||| ||| | | ||| | || | ||||| || | ||||| ..A.GTR.FD..TGTTHRIEDVVDGRK.I.VVA.A.DGTLHAR..VS...QG..DV.G.RIA.GAI.W.T..HKV.T.YG.R..GELRK.DR.A.RDVET --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 -------110-------120-------130--------- GELRYSVIREVLPTRRARTFDLEVEELHTLVAEGVVVHN ||| |||||||| || | | | | ||||||||| GEL.YSVIREVL.TR.A.T.D...E....LVAEGVVVH -------110-------120-------130--------
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 CLAEGTRIFDPVTGTTHRIEDVVDGRKPIHVVAAAKDGTLHARPVVSWFDQGTRDVIGLRIAGGAILWATPDHKVLTEYGWRAAGELRKGDRVAVRDVET ||||||||| |||| ||||||||||| ||||| ||||||||| ||||| ||| || | ||| ||||| | ||||||||||| |||||| || | ||||| .LAEGTRIFD.VTGT.HRIEDVVDGRK.IHVVA.AKDGTLHAR.VVSWF.QGT.DV.G.RIA.GAILW.T.DHKVLTEYGWR.AGELRK.DR.A.RDVET --------10--------20--------30--------40--------50--------60--------70--------80--------90-------100 -------110-------120-------130--------- GELRYSVIREVLPTRRARTFDLEVEELHTLVAEGVVVHN ||| |||| ||| |||||||| ||| | |||||||| GEL.YSVI.EVL.TRRARTFD.EVE.L.TLVAEGVV -------110-------120-------130------