BMRBj - BMREntryMaster

Found 1 Document (0.641 seconds)

BMRB: 30480

6DSL

Consensus engineered intein (Cat) with atypical split site

Authors

Sekar, G., Stevens, A.J., Muir, T.W., Cowburn, D.

Assembly

CatN, CatC

Entity

1. Consensus engineered intein CatN (polymer), 33 monomers, 3773.200 Da Detail

EFEALSGDTM IEILDDDGII QKISMEDLYQ RLA

2. Consensus engineered intein CatC (polymer), 118 monomers, 13499.07 Da Detail

DYKDDDDKMF KLNTKNIKVL TPSGFKSFSG IQKVYKPFYH HIIFDDGSEI KCSDNHSFGK DKIKASTIKV GDYLQGKKVL YNEIVEEGIY LYDLLNVGED NLYYTNGIVS HACESRGK

Total weight

17272.27 Da

Max. entity weight

13499.07 Da

Source organism

Escherichia phage T7

Exptl. method

solution NMR

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 98.7 %, Completeness: 90.5 %, Completeness (bb): 95.8 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	90.5 % (1628 of 1799)	91.8 % (859 of 936)	88.3 % (621 of 703)	92.5 % (148 of 160)
Backbone	95.8 % (864 of 902)	95.5 % (298 of 312)	96.1 % (424 of 441)	95.3 % (142 of 149)
Sidechain	86.7 % (898 of 1036)	89.9 % (561 of 624)	82.5 % (331 of 401)	54.5 % (6 of 11)
Aromatic	55.4 % (92 of 166)	73.5 % (61 of 83)	37.3 % (31 of 83)
Methyl	98.1 % (151 of 154)	97.4 % (75 of 77)	98.7 % (76 of 77)

1. Consensus engineered intein CatN

EFEALSGDTM IEILDDDGII QKISMEDLYQ RLA

2. Consensus engineered intein CatC

DYKDDDDKMF KLNTKNIKVL TPSGFKSFSG IQKVYKPFYH HIIFDDGSEI KCSDNHSFGK DKIKASTIKV GDYLQGKKVL YNEIVEEGIY LYDLLNVGED NLYYTNGIVS HACESRGK

Show sample condition

Sample

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 6.8, Details 300 uM [U-99% 13C; U-99% 15N] CatN, 300 uM [U-99% 13C; U-99% 15N] CatC, 50 mM sodium phosphate, 100 mM sodium chloride, 2 mM TCEP, 90% H2O/10% D2O

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

LACS Plot; CA

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	internal	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	internal	indirect	0.1013291

Referencing offset: -0.13 ppm, Outliers: 1 Detail

LACS Plot; CB

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	internal	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	internal	indirect	0.1013291

Referencing offset: -0.13 ppm, Outliers: 1 Detail

LACS Plot; HA

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	internal	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	internal	indirect	0.1013291

Referencing offset: -0.07 ppm, Outliers: 1 Detail

LACS Plot; CO

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	internal	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	internal	indirect	0.1013291

Referencing offset: 0.67 ppm, Outliers: 3 Detail

Release date

2018-07-05

Citation

An Atypical Mechanism of Split Intein Molecular Recognition and Folding
Stevens, A.J., Sekar, G., Gramespacher, J.A., Cowburn, D., Muir, T.W.
J. Am. Chem. Soc. (2018), 140, 11791-11799, PubMed 30156841 , DOI 10.1021/jacs.8b07334 , Abstract

Related entities 1. Consensus engineered intein CatN, : 1 : 26 entities Detail

Experiments performed 10 experiments Detail

1 3D 1H-13C NOESY aliphatic

Instrument

Bruker AvanceIII - 900 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 6.8, Details 300 uM [U-99% 13C; U-99% 15N] CatN, 300 uM [U-99% 13C; U-99% 15N] CatC, 50 mM sodium phosphate, 100 mM sodium chloride, 2 mM TCEP, 90% H2O/10% D2O

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

2 3D 1H-13C NOESY aromatic

Instrument

Bruker DRX - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

3 3D 1H-15N NOESY

Instrument

Bruker AvanceIII - 900 MHz

Sample

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

4 3D HN(CA)CO

Instrument

Bruker DRX - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

5 3D HNCO

Instrument

Bruker DRX - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

6 3D CBCA(CO)NH

Instrument

Bruker AvanceIII - 800 MHz

Sample

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

7 3D HNCACB

Instrument

Bruker AvanceIII - 800 MHz

Sample

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

8 3D HNCA

Instrument

Bruker AvanceIII - 800 MHz

Sample

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

9 3D HCCH-TOCSY

Instrument

Bruker AvanceIII - 900 MHz

Sample

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

10 3D HCCCONH

Instrument

Bruker DRX - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	CatN	[U-99% 13C; U-99% 15N]	300 uM
2	CatC	[U-99% 13C; U-99% 15N]	300 uM
3	sodium phosphate	natural abundance	50 mM
4	sodium chloride	natural abundance	100 mM
5	TCEP	natural abundance	2 mM

NMR combined restraints 3 contents Detail

Properties

Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints	combined_30480_6dsl.nef
Input source #2: Coordindates	6dsl.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

None

Other bonds (neither disulfide, covalent nor hydrogen bonds, e.g. Zinc–sulphur bond)

None

Non-standard residues

None

Sequence alignments

Entity_assembly_ID (NMR): A vs Auth_asym_ID (model): A

-----------10--------20--------30
EFEALSGDTMIEILDDDGIIQKISMEDLYQRLA
|||||||||||||||||||||||||||||||||
EFEALSGDTMIEILDDDGIIQKISMEDLYQRLA
--------10--------20--------30---

Entity_assembly_ID (NMR): B vs Auth_asym_ID (model): B

------30--------40--------50--------60--------70--------80--------90-------100-------110-------120--
DYKDDDDKMFKLNTKNIKVLTPSGFKSFSGIQKVYKPFYHHIIFDDGSEIKCSDNHSFGKDKIKASTIKVGDYLQGKKVLYNEIVEEGIYLYDLLNVGED
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
DYKDDDDKMFKLNTKNIKVLTPSGFKSFSGIQKVYKPFYHHIIFDDGSEIKCSDNHSFGKDKIKASTIKVGDYLQGKKVLYNEIVEEGIYLYDLLNVGED
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100

-----130-------140
NLYYTNGIVSHACESRGK
||||||||||||||||||
NLYYTNGIVSHACESRGK
-------110--------

Chain assignments

Entity_assembly_ID (NMR)	Auth_asym_ID (model)	Length	Unmapped	Conflict	Sequence coverage (%)
A	A	33	0	0	100.0
B	B	118	0	0	100.0

Content subtype: combined_30480_6dsl.nef

#	Content subtype	Saveframe	Status	# of rows (sets)	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	assigned_chemical_shifts_1	Warning	1614		93.9 (chain: A, length: 33), 99.2 (chain: B, length: 118)
1	Distance restraints	CNS/XPLOR_distance_constraints_2	OK	4054 (1)	noe	90.9 (chain: A, length: 33), 99.2 (chain: B, length: 118)

Assigned chemical shifts

Saveframe: assigned_chemical_shifts_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 33, Sequence coverage: 93.9%
  - Entity_assembly_ID: B, Chain length: 118, Sequence coverage: 99.2%
- Total number of assignments
  - ¹³C chemical shifts: 617
  - ¹H chemical shifts: 852
  - ¹⁵N chemical shifts: 145
- Completeness of assignments
  - Entity_assemble_ID: A
  - Entity_assemble_ID: B
- Redox state of CYS
- Peptide bond of PRO
- Tautomeric state of HIS
- Rotameric state of ILE , LEU, and VAL
- Histogram of normalized assigned chemical shifts
- Random coil index and derived parameters (S² and NMR RMSD)
  - Chain_ID: A
  - Chain_ID: B

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	149	131	87.9
¹H chemical shifts	197	170	86.3
¹⁵N chemical shifts	36	30	83.3

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	66	61	92.4
¹H chemical shifts	68	62	91.2
¹⁵N chemical shifts	33	30	90.9

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	83	70	84.3
¹H chemical shifts	129	108	83.7
¹⁵N chemical shifts	3	0	0.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	23	23	100.0
¹H chemical shifts	23	23	100.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	9	2	22.2
¹H chemical shifts	9	4	44.4

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	554	486	87.7
¹H chemical shifts	739	682	92.3
¹⁵N chemical shifts	126	115	91.3

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	236	228	96.6
¹H chemical shifts	244	235	96.3
¹⁵N chemical shifts	116	110	94.8

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	318	258	81.1
¹H chemical shifts	495	447	90.3
¹⁵N chemical shifts	10	5	50.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	57	56	98.2
¹H chemical shifts	57	56	98.2

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	74	29	39.2
¹H chemical shifts	74	57	77.0

Distance restraints

Saveframe: CNS/XPLOR_distance_constraints_2
- Status: OK
- Experiment type: noe
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 33, Sequence coverage: 90.9%
  - Entity_assembly_ID: B, Chain length: 118, Sequence coverage: 99.2%
- Total number of restraints: 4052
- Weight of restraints: 1.0 (4052)
- Potential type of restraints: square-well-parabolic (4052)
- Range of distance target values: 1.71, 3.88 (Å)
- Histogram of distance target values
- Histogram of discrepancy in distance target values
- Distance restraints per residue
  - Chain_ID: A
  - Chain_ID: B
- Distance restraints on contact map

Keywords Consensus Engineering, Intein Protein, Protein Engineering, SPLICING, Trans Splicing

Search

Query history

Resources

Search for primary databases

Search for subsidiary databases

Display

Hits per page

Statistics

Query Changes will take effect after "OK" button is selected.

Maximum hits

Default conjunction operator

Omni-box

Search filter

Sequence search (BLAST)

E-value

Query sequence coverage

Miscellaneous

The number of previewed publications

Completeness of assigned chemical shift

Sequence chart about top aligned regions

Sequence chart about interacting regions

Hit context with highlight

Link destinations for BMRB, PDB and EMDB

Found 1 Document (0.641 seconds)

BMRB: 30480

Sample

Chemical shift reference

Chemical shift reference

Chemical shift reference

Chemical shift reference

Related entities 1. Consensus engineered intein CatN, : 1 : 26 entities Detail

Experiments performed 10 experiments Detail

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

NMR combined restraints 3 contents Detail