BMRB Entry 26349

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR26349

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Title:
Stereospecific Nuclear Magnetic Resonance Assignments of the Aromatic and Aliphatic Groups of Phenylalanine in the Malate Synthasa G (MSG) by Relaxation Optimized Stereo Array Isotope Labeling
Deposition date:
2024-08-16
Original release date:
2025-08-16
Authors:
Miyanoiri, Yohei; Takeda, Mitsuhiro; Okuma, Kosuke; Terauchi, Tsutomu; Kainosho, Masatsune
Citation:

Citation: Miyanoiri, Yohei; Takeda, Mitsuhiro; Okuma, Kosuke; Terauchi, Tsutomu; Kainosho, Masatsune. "Enhancing solution structural analysis of large molecular proteins through optimal stereo array isotope labeling of aromatic amino acids"  Biophys. Chem. 315, 107328-107328 (2024).
PubMed: 39341158

Assembly members:

Assembly members:
Malate Synthase G, polymer, 731 residues, 81537.83 Da.
Magesium ion, non-polymer, Formula weight is not available

Natural source:

Natural source:   Common Name: not available   Taxonomy ID: 83333   Superkingdom: Bacteria   Kingdom: not available   Genus/species: Escherichia coli

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pET28b

Entity Sequences (FASTA):

Entity Sequences (FASTA):
Malate Synthase G: MAQTITQSRLRIDANFKRFV DEEVLPGTGLDAAAFWRNFD EIVHDLAPENRQLLAERDRI QAALDEWHRSNPGPVKDKAA YKSFLRELGYLVPQPERVTV ETTGIDSEITSQAGPQLVVP AMNARYALNAANARWGSLYD ALYGSDIIPQEGAMVSGYDP QRGEQVIAWVRRFLDESLPL ENGSYQDVVAFKVVDKQLRI QLKNGKETTLRTPAQFVGYR GDAAAPTCILLKNNGLHIEL QIDANGRIGKDDPAHINDVI VEAAISTILDCEDSVAAVDA EDKILLYRNLLGLMQGTLQE KMEKNGRQIVRKLNDDRHYT AADGSEISLHGRSLLFIRNV GHLMTIPVIWDSEGNEIPEG ILDGVMTGAIALYDLKVQKN SRTGSVYIVKPKMHGPQEVA FANKLFTRIETMLGMAPNTL KMGIMDEERRTSLNLRSCIA QARNRVAFINTGFLDRTGDE MHSVMEAGPMLRKNQMKSTP WIKAYERNNVLSGLFCGLRG KAQIGKGMWAMPDLMADMYS QKGDQLRAGANTAWVPSPTA ATLHALHYHQTNVQSVQANI AQTEFNAEFEPLLDDLLTIP VAENANWSAQEIQQELDNNV QGILGYVVRWVEQGIGCSKV PDIHNVALMEDRATLRISSQ HIANWLRHGILTKEQVQASL ENMAKVVDQQNAGDPAYRPM AGNFANSCAFKAASDLIFLG VKQPNGYTEPLLHAWRLREK ESHLEHHHHHH

Data sets:
Data typeCount
13C chemical shifts76
1H chemical shifts95

Additional metadata:

  • Assembly
  • Samples and Experiments
  • Software
  • Spectrometers
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Assembly:

Entity Assembly IDEntity NameEntity ID
111
222

Entities:

Entity 1, 1 731 residues - 81537.83 Da.

Residues 726-731 represent a non-native affinity tag.

1   METALAGLNTHRILETHRGLNSERARGLEU
2   ARGILEASPALAASNPHELYSARGPHEVAL
3   ASPGLUGLUVALLEUPROGLYTHRGLYLEU
4   ASPALAALAALAPHETRPARGASNPHEASP
5   GLUILEVALHISASPLEUALAPROGLUASN
6   ARGGLNLEULEUALAGLUARGASPARGILE
7   GLNALAALALEUASPGLUTRPHISARGSER
8   ASNPROGLYPROVALLYSASPLYSALAALA
9   TYRLYSSERPHELEUARGGLULEUGLYTYR
10   LEUVALPROGLNPROGLUARGVALTHRVAL
11   GLUTHRTHRGLYILEASPSERGLUILETHR
12   SERGLNALAGLYPROGLNLEUVALVALPRO
13   ALAMETASNALAARGTYRALALEUASNALA
14   ALAASNALAARGTRPGLYSERLEUTYRASP
15   ALALEUTYRGLYSERASPILEILEPROGLN
16   GLUGLYALAMETVALSERGLYTYRASPPRO
17   GLNARGGLYGLUGLNVALILEALATRPVAL
18   ARGARGPHELEUASPGLUSERLEUPROLEU
19   GLUASNGLYSERTYRGLNASPVALVALALA
20   PHELYSVALVALASPLYSGLNLEUARGILE
21   GLNLEULYSASNGLYLYSGLUTHRTHRLEU
22   ARGTHRPROALAGLNPHEVALGLYTYRARG
23   GLYASPALAALAALAPROTHRCYSILELEU
24   LEULYSASNASNGLYLEUHISILEGLULEU
25   GLNILEASPALAASNGLYARGILEGLYLYS
26   ASPASPPROALAHISILEASNASPVALILE
27   VALGLUALAALAILESERTHRILELEUASP
28   CYSGLUASPSERVALALAALAVALASPALA
29   GLUASPLYSILELEULEUTYRARGASNLEU
30   LEUGLYLEUMETGLNGLYTHRLEUGLNGLU
31   LYSMETGLULYSASNGLYARGGLNILEVAL
32   ARGLYSLEUASNASPASPARGHISTYRTHR
33   ALAALAASPGLYSERGLUILESERLEUHIS
34   GLYARGSERLEULEUPHEILEARGASNVAL
35   GLYHISLEUMETTHRILEPROVALILETRP
36   ASPSERGLUGLYASNGLUILEPROGLUGLY
37   ILELEUASPGLYVALMETTHRGLYALAILE
38   ALALEUTYRASPLEULYSVALGLNLYSASN
39   SERARGTHRGLYSERVALTYRILEVALLYS
40   PROLYSMETHISGLYPROGLNGLUVALALA
41   PHEALAASNLYSLEUPHETHRARGILEGLU
42   THRMETLEUGLYMETALAPROASNTHRLEU
43   LYSMETGLYILEMETASPGLUGLUARGARG
44   THRSERLEUASNLEUARGSERCYSILEALA
45   GLNALAARGASNARGVALALAPHEILEASN
46   THRGLYPHELEUASPARGTHRGLYASPGLU
47   METHISSERVALMETGLUALAGLYPROMET
48   LEUARGLYSASNGLNMETLYSSERTHRPRO
49   TRPILELYSALATYRGLUARGASNASNVAL
50   LEUSERGLYLEUPHECYSGLYLEUARGGLY
51   LYSALAGLNILEGLYLYSGLYMETTRPALA
52   METPROASPLEUMETALAASPMETTYRSER
53   GLNLYSGLYASPGLNLEUARGALAGLYALA
54   ASNTHRALATRPVALPROSERPROTHRALA
55   ALATHRLEUHISALALEUHISTYRHISGLN
56   THRASNVALGLNSERVALGLNALAASNILE
57   ALAGLNTHRGLUPHEASNALAGLUPHEGLU
58   PROLEULEUASPASPLEULEUTHRILEPRO
59   VALALAGLUASNALAASNTRPSERALAGLN
60   GLUILEGLNGLNGLULEUASPASNASNVAL
61   GLNGLYILELEUGLYTYRVALVALARGTRP
62   VALGLUGLNGLYILEGLYCYSSERLYSVAL
63   PROASPILEHISASNVALALALEUMETGLU
64   ASPARGALATHRLEUARGILESERSERGLN
65   HISILEALAASNTRPLEUARGHISGLYILE
66   LEUTHRLYSGLUGLNVALGLNALASERLEU
67   GLUASNMETALALYSVALVALASPGLNGLN
68   ASNALAGLYASPPROALATYRARGPROMET
69   ALAGLYASNPHEALAASNSERCYSALAPHE
70   LYSALAALASERASPLEUILEPHELEUGLY
71   VALLYSGLNPROASNGLYTYRTHRGLUPRO
72   LEULEUHISALATRPARGLEUARGGLULYS
73   GLUSERHISLEUGLUHISHISHISHISHIS
74   HIS

Entity 2, 2 - Formula weight is not available

1   MG

Samples:

sample_1: Malate Synthase G, [RO-SAIL_13CHd,b3 Phe; U-2H; 15N], 0.3 mM; sodium azide 1 mM; DL-1,4-DTT 5 mM; magnesium chloride 20 mM; sodium phosphate 20 mM; DSS 0.01 % w/v; D2O, [U-2H], 5 % v/v; H2O 95%

sample_2: Malate Synthase G, [RO-SAIL_13CHd,b2 Phe; U-2H; 15N], 0.3 mM; sodium azide 1 mM; DL-1,4-DTT 5 mM; magnesium chloride 20 mM; sodium phosphate 20 mM; DSS 0.01 % w/v; D2O, [U-2H], 5 % v/v; H2O 95%

sample_3: Malate Synthase G, [RO-SAIL_13CHb3 Phe; U-2H; 15N], 0.3 mM; sodium azide 1 mM; DL-1,4-DTT 5 mM; magnesium chloride 20 mM; sodium phosphate 20 mM; DSS 0.01 % w/v; D2O, [U-2H], 5 % v/v; H2O 95%

sample_4: Malate Synthase G, [RO-SAIL_13CHb2 Phe; U-2H; 15N], 0.3 mM; sodium azide 1 mM; DL-1,4-DTT 5 mM; magnesium chloride 20 mM; sodium phosphate 20 mM; DSS 0.01 % w/v; D2O, [U-2H], 5 % v/v; H2O 95%

sample_6: Malate Synthase G, [RO-SAIL_13CHz,d,b2 Phe; U-2H; 15N], 0.3 mM; sodium azide 1 mM; DL-1,4-DTT 5 mM; magnesium chloride 20 mM; sodium phosphate 20 mM; DSS 0.01 % w/v; D2O, [U-2H], 5 % v/v; H2O 95%

sample_7: Malate Synthase G, [RO-SAIL_Hz,d,b2 Phe; U-2H; 15N], 0.3 mM; sodium azide 1 mM; DL-1,4-DTT 5 mM; magnesium chloride 20 mM; sodium phosphate 20 mM; DSS 0.01 % w/v; D2O, [U-2H], 99 % v/v

sample_conditions_1: ionic strength: 0.02 M; pH: 7.1; pressure: 1 atm; temperature: 310 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-13C TROSYsample_1isotropicsample_conditions_1
2D 1H-13C TROSYsample_2isotropicsample_conditions_1
2D 1H-13C TROSYsample_3isotropicsample_conditions_1
2D 1H-13C TROSYsample_4isotropicsample_conditions_1
3D 1H-13C NOESY aliphaticsample_1isotropicsample_conditions_1
3D 1H-13C NOESY aliphaticsample_2isotropicsample_conditions_1
3D 1H-13C NOESY aromaticsample_6isotropicsample_conditions_1
1D 1Hsample_7isotropicsample_conditions_1
2D 1H-1H NOESYsample_7isotropicsample_conditions_1

Software:

NMRFAM-SPARKY v1.47 - chemical shift assignment, peak picking

TOPSPIN v3.6.2 - data analysis, processing

NMR spectrometers:

  • Bruker AVANCE III 950 MHz
  • Bruker AVANCE III 900 MHz
  • Bruker AVANCE III HD 800 MHz

Related Database Links:

PDB
EMBL CAA52639
REF NP_417450