BMRB Entry 50548

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR50548

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Title:
1H, 13C, and 15N NMR chemical shift assignment of the complex formed by the first EPEC EspF repeat and N-WASP GTPase binding domain
Deposition date:
2020-11-06
Original release date:
2021-03-15
Authors:
Karjalainen, Mikael; Hellman, Maarit; Tossavainen, Helena; Permi, Perttu
Citation:

Citation: Karjalainen, Mikael; Hellman, Maarit; Tossavainen, Helena; Permi, Perttu. "1H, 13C, and 15N NMR chemical shift assignment of the complex formed by the first EPEC EspF repeat and N-WASP GTPase binding domain"  Biomol. NMR Assignments 15, 213-217 (2021).
PubMed: 33475933

Assembly members:

Assembly members:
entity_1, polymer, 48 residues, 4909.54 Da.
entity_2, polymer, 65 residues, 7254.04 Da.

Natural source:

Natural source:   Common Name: E. coli   Taxonomy ID: 562   Superkingdom: Bacteria   Kingdom: not available   Genus/species: Escherichia coli

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pET15b

Entity Sequences (FASTA):

Entity Sequences (FASTA):
entity_1: GPSRPAPPPPTSGQASGASR PLPPIAQALKDHLAAYELSK ASETVNFK
entity_2: GSNFQHIGHVGWDPNTGFDL NNLDPELKNLFDMCGISEAQ LKDRETSKVIYDFIEKTGGV EAVKN

Data sets:
Data typeCount
13C chemical shifts381
15N chemical shifts114
1H chemical shifts720

Additional metadata:

  • Assembly
  • Samples and Experiments
  • Software
  • Spectrometers
  • Hide all

Assembly:

Entity Assembly IDEntity NameEntity ID
1EspF repeat1
2N-WASP GBD2

Entities:

Entity 1, EspF repeat 48 residues - 4909.54 Da.

This is the first proline-rich repeat of the LEE-encoded effector EspF (EspF) protein from enteropathogenic Escherichia coli (EPEC). N-terminal glycine residue 72 is a cloning artifact. Numbering scheme for residues 73-119 follows numbering from UniProtKB entry B7UM88.

1   GLYPROSERARGPROALAPROPROPROPRO
2   THRSERGLYGLNALASERGLYALASERARG
3   PROLEUPROPROILEALAGLNALALEULYS
4   ASPHISLEUALAALATYRGLULEUSERLYS
5   ALASERGLUTHRVALASNPHELYS

Entity 2, N-WASP GBD 65 residues - 7254.04 Da.

This is the GTPase binding domain (GBD) of the Neural Wiskott-Aldrich syndrome protein (N-WASP) from Homo sapiens. N-terminal glycine residue 206 is a cloning artifact. Numbering scheme for residues 207-270 follows numbering from UniProtKB entry O00401.

1   GLYSERASNPHEGLNHISILEGLYHISVAL
2   GLYTRPASPPROASNTHRGLYPHEASPLEU
3   ASNASNLEUASPPROGLULEULYSASNLEU
4   PHEASPMETCYSGLYILESERGLUALAGLN
5   LEULYSASPARGGLUTHRSERLYSVALILE
6   TYRASPPHEILEGLULYSTHRGLYGLYVAL
7   GLUALAVALLYSASN

Samples:

sample_1: LEE-encoded effector EspF (EspF), [U-13C; U-15N], 0.3 mM; Neural Wiskott-Aldrich syndrome protein (N-WASP) GTPase binding domain (GBD) 0.36 mM; sodium phosphate 20 mM; sodium chloride 50 mM

sample_2: LEE-encoded effector EspF (EspF), [U-13C; U-15N], 0.45 mM; Neural Wiskott-Aldrich syndrome protein (N-WASP) GTPase binding domain (GBD) 0.5 mM; sodium phosphate 20 mM; sodium chloride 50 mM

sample_3: Neural Wiskott-Aldrich syndrome protein (N-WASP) GTPase binding domain (GBD), [U-13C; U-15N], 0.5 mM; LEE-encoded effector EspF (EspF) 0.6 mM; sodium phosphate 20 mM; sodium chloride 50 mM

sample_4: Neural Wiskott-Aldrich syndrome protein (N-WASP) GTPase binding domain (GBD), [U-13C; U-15N], 0.5 mM; LEE-encoded effector EspF (EspF) 1.0 mM; sodium phosphate 20 mM; sodium chloride 50 mM

sample_conditions_1: pH: 7; temperature: 303 K

sample_conditions_2: pH: 6.5; temperature: 298 K

sample_conditions_3: pH: 6.5; temperature: 298 K

sample_conditions_4: pH: 6.5; temperature: 298 K

Experiments:

NameSampleSample stateSample conditions
2D CONsample_1isotropicsample_conditions_1
2D 1H-15N HSQCsample_2isotropicsample_conditions_2
3D 1H-15N NOESYsample_2isotropicsample_conditions_2
3D 1H-13C NOESYsample_2isotropicsample_conditions_2
3D HNCACBsample_2isotropicsample_conditions_2
3D HN(CO)CACBsample_2isotropicsample_conditions_2
3D (H)CC(CO)NHsample_2isotropicsample_conditions_2
3D H(CC)(CO)NHsample_2isotropicsample_conditions_2
3D HC(C)H-COSYsample_2isotropicsample_conditions_2
2D (HB)CB(CGCD)HDsample_2isotropicsample_conditions_2
2D (HB)CB(CGCDCE)HEsample_2isotropicsample_conditions_2
2D 1H-13C HSQC aromaticsample_2isotropicsample_conditions_2
2D 1H-13C HSQC aliphaticsample_2isotropicsample_conditions_2
3D HBHA(CO)NHsample_2isotropicsample_conditions_2
3D 1H-13C NOESY aromaticsample_2isotropicsample_conditions_2
2D 1H-15N HSQCsample_3isotropicsample_conditions_3
3D HNCACBsample_3isotropicsample_conditions_3
3D HN(CO)CACBsample_3isotropicsample_conditions_3
3D 1H-15N NOESYsample_3isotropicsample_conditions_3
3D (H)CC(CO)NHsample_3isotropicsample_conditions_3
3D H(CC)(CO)NHsample_3isotropicsample_conditions_3
3D HC(C)H-COSYsample_3isotropicsample_conditions_3
2D (HB)CB(CGCD)HDsample_3isotropicsample_conditions_3
2D (HB)CB(CGCDCE)HEsample_3isotropicsample_conditions_3
2D 1H-13C HSQC aromaticsample_3isotropicsample_conditions_3
3D HBHA(CO)NHsample_3isotropicsample_conditions_3
2D CONsample_3isotropicsample_conditions_3
3D 1H-13C NOESYsample_4isotropicsample_conditions_4
3D 1H-13C NOESY aromaticsample_4isotropicsample_conditions_4
2D 1H-13C HSQC aliphaticsample_4isotropicsample_conditions_4
3D (H)CCH3-TOCSYsample_4isotropicsample_conditions_4
3D HC(C)H-COSYsample_4isotropicsample_conditions_4
4D (HACA)CONCAHAsample_1isotropicsample_conditions_1

Software:

TOPSPIN v3.5 - collection, processing

ANALYSIS v2.4.2 - chemical shift assignment

NMR spectrometers:

  • Bruker AVANCE III 800 MHz

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