BMRB Entry 26981

Name: Fused Npu DnaE GEP loop mutant from Nostoc punctiforme with Phe +2 Extein
Published: 2017-08-18

Chem Shift validation: AVS_full, LACS
BMRB Entry DOI: doi:10.13018/BMR26981

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Nicholas Rego and David Koes
3Dmol.js: molecular visualization with WebGL
Bioinformatics (2015) 31 (8): 1322-1324 doi:10.1093/bioinformatics/btu829

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Title: Fused Npu DnaE GEP loop mutant from Nostoc punctiforme with Phe +2 Extein PubMed: 28739907

Deposition date: 2016-12-21 Original release date: 2017-08-18

Authors: Sekar, Giridhar; Cowburn, David

Citation: Stevens, Adam; Sekar, Giridhar; Shah, Neel; Mostafavi, Ana; Cowburn, David; Muir, Tom. "A promiscuous split intein with expanded protein engineering applications" Proc. Natl. Acad. Sci. U. S. A. 114, 8538-8543 (2017).

Assembly members:
Npu_DnaE, polymer, 144 residues, 16493.6656 Da.

Natural source: Common Name: Nostoc punctiforme Taxonomy ID: 272131 Superkingdom: Bacteria Kingdom: not available Genus/species: Nostoc punctiforme

Experimental source: Production method: recombinant technology Host organism: Escherichia coli Vector: pET

Entity Sequences (FASTA):
Npu_DnaE: AEYALSYETEILTVEYGLLP IGKIVEKRIECTVYSVDNNG NIYTQPVAQWHDRGEQEVFE YCLEDGSLIRATKDHKFMTV DGQMLPIDEIFERELDLMRV DNLPNMIKIATRKYLGKQNV YDIGVGEPHNFALKNGFIAS ACFN

Data sets:

assigned_chemical_shifts
- assigned_chem_shift_list

Data type	Count
13C chemical shifts	417
15N chemical shifts	142
1H chemical shifts	149

Additional metadata:

Assembly
Samples and Experiments
Software
Spectrometers
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Assembly:

Entity Assembly ID	Entity Name	Entity ID
1	Fused Npu DnaE GEP loop mutant	1

Entities:

Entity 1, Fused Npu DnaE GEP loop mutant 144 residues - 16493.6656 Da.

1

ALA

GLU

TYR

ALA

LEU

SER

TYR

GLU

THR

GLU

2

ILE

LEU

THR

VAL

GLU

TYR

GLY

LEU

PRO

3

ILE

GLY

LYS

ILE

VAL

GLU

LYS

ARG

ILE

GLU

4

CYS

THR

VAL

TYR

SER

VAL

ASP

ASN

GLY

5

ASN

ILE

TYR

THR

GLN

PRO

VAL

ALA

GLN

TRP

6

HIS

ASP

ARG

GLY

GLU

GLN

GLU

VAL

PHE

GLU

7

TYR

CYS

LEU

GLU

ASP

GLY

SER

LEU

ILE

ARG

8

ALA

THR

LYS

ASP

HIS

LYS

PHE

MET

THR

VAL

9

ASP

GLY

GLN

MET

LEU

PRO

ILE

ASP

GLU

ILE

10

PHE

GLU

ARG

GLU

LEU

ASP

LEU

MET

ARG

VAL

11

ASP

ASN

LEU

PRO

ASN

MET

ILE

LYS

ILE

ALA

12

THR

ARG

LYS

TYR

LEU

GLY

LYS

GLN

ASN

VAL

13

TYR

ASP

ILE

GLY

VAL

GLY

GLU

PRO

HIS

ASN

14

PHE

ALA

LEU

LYS

ASN

GLY

PHE

ILE

ALA

SER

15

ALA

CYS

PHE

ASN

Samples:

sample_1: sodium phosphate 20 mM; TCEP 2 mM; Npu DnaE, [U-99% 13C; U-99% 15N], 400 mM

CondSet1: pH: 6.0; pressure: 1 atm; temperature: 300 K

Experiments:

Name	Sample	Sample state	Sample conditions
2D 1H-15N HSQC	sample_1	isotropic	CondSet1
3D HNCACB	sample_1	isotropic	CondSet1
3D CBCA(CO)NH	sample_1	isotropic	CondSet1
3D HN(CA)CO	sample_1	isotropic	CondSet1
3D HNCO	sample_1	isotropic	CondSet1

Software:

CcpNmr_Analysis v2.4, CCPN - Spectrum processing, chemical shift assignment

NMR spectrometers:

Bruker Avance 900 MHz
Bruker Avance 600 MHz

Download HSQC peak lists in one of the following formats:
CSV: Backbone or all simulated shifts
SPARKY: Backbone or all simulated shifts