BMRB Entry 50530

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR50530

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Title:
Methyl assignments of recombinant wild-type human alpha1-antitrypsin
Deposition date:
2020-10-28
Original release date:
2021-04-22
Authors:
Jagger, Alistair; Waudby, Christopher; Irving, James; Christodoulou, John; Lomas, David
Citation:

Citation: Jagger, Alistair; Waudby, Christopher; Irving, James; Christodoulou, John; Lomas, David. "High-resolution ex vivo NMR spectroscopy of human Z a1-antitrypsin"  Nat. Commun. 11, 6371-6371 (2020).
PubMed: 33311470

Assembly members:

Assembly members:
entity_1, polymer, 404 residues, Formula weight is not available

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: pQE-30

Entity Sequences (FASTA):

Entity Sequences (FASTA):
entity_1: MRGSHHHHHHTDPQGDAAQK TDTSHHDQDHPTFNKITPNL AEFAFSLYRQLAHQSNSTNI FFSPVSIATAFAMLSLGTKA DTHDEILEGLNFNLTEIPEA QIHEGFQELLRTLNQPDSQL QLTTGNGLFLSEGLKLVDKF LEDVKKLYHSEAFTVNFGDT EEAKKQINDYVEKGTQGKIV DLVKELDRDTVFALVNYIFF KGKWERPFEVKDTEEEDFHV DQVTTVKVPMMKRLGMFNIQ HCKKLSSWVLLMKYLGNATA IFFLPDEGKLQHLENELTHD IITKFLENEDRRSASLHLPK LSITGTYDLKSVLGQLGITK VFSNGADLSGVTEEAPLKLS KAVHKAVLTIDEKGTEAAGA MFLEAIPMSIPPEVKFNKPF VFLMIEQNTKSPLFMGKVVN PTQK

Data sets:
Data typeCount
13C chemical shifts196
1H chemical shifts588

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1AAT1

Entities:

Entity 1, AAT 404 residues - Formula weight is not available

Residues 1-11 represent a non-native affinity tag. Subsequent residue numbers are offset by 10 from that of the mature protein.

1   METARGGLYSERHISHISHISHISHISHIS
2   THRASPPROGLNGLYASPALAALAGLNLYS
3   THRASPTHRSERHISHISASPGLNASPHIS
4   PROTHRPHEASNLYSILETHRPROASNLEU
5   ALAGLUPHEALAPHESERLEUTYRARGGLN
6   LEUALAHISGLNSERASNSERTHRASNILE
7   PHEPHESERPROVALSERILEALATHRALA
8   PHEALAMETLEUSERLEUGLYTHRLYSALA
9   ASPTHRHISASPGLUILELEUGLUGLYLEU
10   ASNPHEASNLEUTHRGLUILEPROGLUALA
11   GLNILEHISGLUGLYPHEGLNGLULEULEU
12   ARGTHRLEUASNGLNPROASPSERGLNLEU
13   GLNLEUTHRTHRGLYASNGLYLEUPHELEU
14   SERGLUGLYLEULYSLEUVALASPLYSPHE
15   LEUGLUASPVALLYSLYSLEUTYRHISSER
16   GLUALAPHETHRVALASNPHEGLYASPTHR
17   GLUGLUALALYSLYSGLNILEASNASPTYR
18   VALGLULYSGLYTHRGLNGLYLYSILEVAL
19   ASPLEUVALLYSGLULEUASPARGASPTHR
20   VALPHEALALEUVALASNTYRILEPHEPHE
21   LYSGLYLYSTRPGLUARGPROPHEGLUVAL
22   LYSASPTHRGLUGLUGLUASPPHEHISVAL
23   ASPGLNVALTHRTHRVALLYSVALPROMET
24   METLYSARGLEUGLYMETPHEASNILEGLN
25   HISCYSLYSLYSLEUSERSERTRPVALLEU
26   LEUMETLYSTYRLEUGLYASNALATHRALA
27   ILEPHEPHELEUPROASPGLUGLYLYSLEU
28   GLNHISLEUGLUASNGLULEUTHRHISASP
29   ILEILETHRLYSPHELEUGLUASNGLUASP
30   ARGARGSERALASERLEUHISLEUPROLYS
31   LEUSERILETHRGLYTHRTYRASPLEULYS
32   SERVALLEUGLYGLNLEUGLYILETHRLYS
33   VALPHESERASNGLYALAASPLEUSERGLY
34   VALTHRGLUGLUALAPROLEULYSLEUSER
35   LYSALAVALHISLYSALAVALLEUTHRILE
36   ASPGLULYSGLYTHRGLUALAALAGLYALA
37   METPHELEUGLUALAILEPROMETSERILE
38   PROPROGLUVALLYSPHEASNLYSPROPHE
39   VALPHELEUMETILEGLUGLNASNTHRLYS
40   SERPROLEUPHEMETGLYLYSVALVALASN
41   PROTHRGLNLYS

Samples:

sample_1: alpha1-antitrypsin, [U-100% 2H; U-100% 15N; 13CH3-100% A(CB)I(CD1)L(CD2)M(CE)V(CG2)], 160 uM; sodium phosphate 25 mM; sodium chloride 50 mM; DTT 1 mM; EDTA 1 mM; sodium azide 0.02 % w/v

sample_2: alpha1-antitrypsin, [U-100% 2H; U-100% 15N; 13CH3-100% I(CD1)L(CD1/CD2)V(CG1/CG2)], 150 uM; sodium phosphate 25 mM; sodium chloride 50 mM; DTT 1 mM; EDTA 1 mM; sodium azide 0.02 % w/v

sample_3: alpha1-antitrypsin, [U-100% 2H; U-100% 15N; 13CH3-100% A(CB)I(CD1/CG2)L(CD1/CD2)V(CG1/CG2)], 165 uM; sodium phosphate 25 mM; sodium chloride 50 mM; DTT 1 mM; EDTA 1 mM; sodium azide 0.02 % w/v

sample_conditions_1: ionic strength: 400 mM; pH*: 7.6; pressure: 1 atm; temperature: 298 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-13C HMQCsample_1isotropicsample_conditions_1
4D CC NOESYsample_1isotropicsample_conditions_1
2D 1H-13C HMQCsample_2isotropicsample_conditions_1
4D CC NOESYsample_2isotropicsample_conditions_1
2D 1H-13C HSQCsample_3isotropicsample_conditions_1
3D HMCMCGsample_3isotropicsample_conditions_1
3D HMCM(CG)CBsample_3isotropicsample_conditions_1
3D HMCM(CGCB)CAsample_3isotropicsample_conditions_1
3D HMCM(CG)COsample_3isotropicsample_conditions_1
3D HMCM(CGCB)COsample_3isotropicsample_conditions_1
3D HMCM(CGCBCA)COsample_3isotropicsample_conditions_1

Software:

NMRPipe - processing

TOPSPIN - collection

CcpNMR - chemical shift assignment, peak picking

MATLAB - peak picking

SMILE - processing

NMR spectrometers:

  • Bruker AVANCE III 900 MHz
  • Bruker AVANCE III 600 MHz

Related Database Links:

BMRB 17804