BMRB Entry 51917

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR51917

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Title:
NT8-13 & ML314 bound enNTS1DM4 with beta-Arrestin-1-3A & PIP2
Deposition date:
2023-04-17
Original release date:
2023-05-04
Authors:
Bumbak, Fabian; Ziarek, Joshua; Robson, Scott; Wu, Hongwei
Citation:

Citation: Bumbak, Fabian; Bower, James; Zemmer, Skylar; Inoue, Asuka; Pons, Miquel; Paniagua, Juan Carlos; Yan, Fei; Ford, James; Wu, Hongwei; Robson, Scott; Bathgate, Ross; Scott, Daniel; Gooley, Paul; Ziarek, Joshua. "Stabilization of pre-existing neurotensin receptor conformational states by b-arrestin-1 and the biased allosteric modulator ML314"  Nat. commun. 14, 3328-3328 (2023).
PubMed: 37286565

Assembly members:

Assembly members:
entity_1, polymer, 404 residues, 44889.10 Da.
entity_2, polymer, 6 residues, 817 Da.
entity_3, non-polymer, Formula weight is not available
entity_4, polymer, 420 residues, 46991.36 Da.
entity_5, non-polymer, Formula weight is not available

Natural source:

Natural source:   Common Name: Rat   Taxonomy ID: 10116   Superkingdom: Eukaryota   Kingdom: Metazoa   Genus/species: Rattus norvegicus

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pDS170

Entity Sequences (FASTA):

Entity Sequences (FASTA):
entity_1: GPGSTSESDTAGPNSDLDVN TDIYSKVLVTAIYLALFVVG TVGNGVTLFTLARKKSLQSL QSRVDYYLGSLALSSLLILL FALPVDVYNFIWVHHPWAFG DAGCKGYYFLREACTYATAL NVVSLSVERYLAICHPFKAK TLLSRSRTKKFISAIWLASA LLSLPMLFTMGLQNLSGDGT HPGGLVCTPIVDTATLRVVI QLNTFMSFLFPMLVASILNT VIARRLTVLVHQAAEQARVS TVGTHNGLEHSTFNVTIEPG RVQALRRGVLVLRAVVIAFV VCWLPYHVRRLMFVYISDEQ WTTALFDFYHYFYMLSNALV YVSAAINPILYNLVSANFRQ VFLSTLASLSPGWRHRRKKR PTFSRKPNSVSSNHAFSTAS GLNDIFEAQKIEWHEGSGLE VLFQ
entity_2: RRPYIL
entity_4: GPSGDKGTRVFKKASPNGKL TVYLGKRDFVDHIDLVDPVD GVVLVDPEYLKERRVYVTLT VAFRYGREDLDVLGLTFRKD LFVANVQSFPPAPEDKKPLT RLQERLIKKLGEHAYPFTFE IPPNLPSSVTLQPGPEDTGK ALGVDYEVKAFVAENLEEKI HKRNSVRLVIRKVQYAPERP GPQPTAETTRQFLMSDKPLH LEASLDKEIYYHGEPISVNV HVTNNTNKTVKKIKISVRQY ADIVLFNTAQYKVPVAMEEA DDTVAPSSTFSKVYTLTPFL ANNREKRGLALDGKLKHEDT NLASSTLLREGANREILGII VSYKVKVKLVVSRGGLLGDL ASSDVAVELPFTLMHPKPKE EPPHREVPENETPVDTNLIE LDTNDDDAAAEDFARQRLKG MKDDKEEEEDGTGSPQLNNR

Data sets:
Data typeCount
13C chemical shifts6
1H chemical shifts18

Additional metadata:

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

Entity Assembly IDEntity NameEntity ID
1enNTS1DM4, conformer A1
2enNTS1DM4, conformer B1
3enNTS1DM4, conformer C1
4NT8-132
5ML3143
6bArr1-3A4
7PIP25

Entities:

Entity 1, enNTS1DM4, conformer A 404 residues - 44889.10 Da.

Thermostabilized rat neurotensin receptor 1 variant starts with the first T (T43) and ends with T416 (truncated). The protein contains a C-terminal Avi-tag and remnants of a 3C-protease cleavage tag (ASGLNDIFEAQKIEWHEGSGLEVLFQ). The N-terminal four residues (GPGS) are remnants of a 3C-protease cleavage site and a short, flexible GS sequence.

1   GLYPROGLYSERTHRSERGLUSERASPTHR
2   ALAGLYPROASNSERASPLEUASPVALASN
3   THRASPILETYRSERLYSVALLEUVALTHR
4   ALAILETYRLEUALALEUPHEVALVALGLY
5   THRVALGLYASNGLYVALTHRLEUPHETHR
6   LEUALAARGLYSLYSSERLEUGLNSERLEU
7   GLNSERARGVALASPTYRTYRLEUGLYSER
8   LEUALALEUSERSERLEULEUILELEULEU
9   PHEALALEUPROVALASPVALTYRASNPHE
10   ILETRPVALHISHISPROTRPALAPHEGLY
11   ASPALAGLYCYSLYSGLYTYRTYRPHELEU
12   ARGGLUALACYSTHRTYRALATHRALALEU
13   ASNVALVALSERLEUSERVALGLUARGTYR
14   LEUALAILECYSHISPROPHELYSALALYS
15   THRLEULEUSERARGSERARGTHRLYSLYS
16   PHEILESERALAILETRPLEUALASERALA
17   LEULEUSERLEUPROMETLEUPHETHRMET
18   GLYLEUGLNASNLEUSERGLYASPGLYTHR
19   HISPROGLYGLYLEUVALCYSTHRPROILE
20   VALASPTHRALATHRLEUARGVALVALILE
21   GLNLEUASNTHRPHEMETSERPHELEUPHE
22   PROMETLEUVALALASERILELEUASNTHR
23   VALILEALAARGARGLEUTHRVALLEUVAL
24   HISGLNALAALAGLUGLNALAARGVALSER
25   THRVALGLYTHRHISASNGLYLEUGLUHIS
26   SERTHRPHEASNVALTHRILEGLUPROGLY
27   ARGVALGLNALALEUARGARGGLYVALLEU
28   VALLEUARGALAVALVALILEALAPHEVAL
29   VALCYSTRPLEUPROTYRHISVALARGARG
30   LEUMETPHEVALTYRILESERASPGLUGLN
31   TRPTHRTHRALALEUPHEASPPHETYRHIS
32   TYRPHETYRMETLEUSERASNALALEUVAL
33   TYRVALSERALAALAILEASNPROILELEU
34   TYRASNLEUVALSERALAASNPHEARGGLN
35   VALPHELEUSERTHRLEUALASERLEUSER
36   PROGLYTRPARGHISARGARGLYSLYSARG
37   PROTHRPHESERARGLYSPROASNSERVAL
38   SERSERASNHISALAPHESERTHRALASER
39   GLYLEUASNASPILEPHEGLUALAGLNLYS
40   ILEGLUTRPHISGLUGLYSERGLYLEUGLU
41   VALLEUPHEGLN

Entity 2, NT8-13 6 residues - 817 Da.

1   ARGARGPROTYRILELEU

Entity 3, ML314 - Formula weight is not available

Entity 4, bArr1-3A 420 residues - 46991.36 Da.

1   GLYPROSERGLYASPLYSGLYTHRARGVAL
2   PHELYSLYSALASERPROASNGLYLYSLEU
3   THRVALTYRLEUGLYLYSARGASPPHEVAL
4   ASPHISILEASPLEUVALASPPROVALASP
5   GLYVALVALLEUVALASPPROGLUTYRLEU
6   LYSGLUARGARGVALTYRVALTHRLEUTHR
7   VALALAPHEARGTYRGLYARGGLUASPLEU
8   ASPVALLEUGLYLEUTHRPHEARGLYSASP
9   LEUPHEVALALAASNVALGLNSERPHEPRO
10   PROALAPROGLUASPLYSLYSPROLEUTHR
11   ARGLEUGLNGLUARGLEUILELYSLYSLEU
12   GLYGLUHISALATYRPROPHETHRPHEGLU
13   ILEPROPROASNLEUPROSERSERVALTHR
14   LEUGLNPROGLYPROGLUASPTHRGLYLYS
15   ALALEUGLYVALASPTYRGLUVALLYSALA
16   PHEVALALAGLUASNLEUGLUGLULYSILE
17   HISLYSARGASNSERVALARGLEUVALILE
18   ARGLYSVALGLNTYRALAPROGLUARGPRO
19   GLYPROGLNPROTHRALAGLUTHRTHRARG
20   GLNPHELEUMETSERASPLYSPROLEUHIS
21   LEUGLUALASERLEUASPLYSGLUILETYR
22   TYRHISGLYGLUPROILESERVALASNVAL
23   HISVALTHRASNASNTHRASNLYSTHRVAL
24   LYSLYSILELYSILESERVALARGGLNTYR
25   ALAASPILEVALLEUPHEASNTHRALAGLN
26   TYRLYSVALPROVALALAMETGLUGLUALA
27   ASPASPTHRVALALAPROSERSERTHRPHE
28   SERLYSVALTYRTHRLEUTHRPROPHELEU
29   ALAASNASNARGGLULYSARGGLYLEUALA
30   LEUASPGLYLYSLEULYSHISGLUASPTHR
31   ASNLEUALASERSERTHRLEULEUARGGLU
32   GLYALAASNARGGLUILELEUGLYILEILE
33   VALSERTYRLYSVALLYSVALLYSLEUVAL
34   VALSERARGGLYGLYLEULEUGLYASPLEU
35   ALASERSERASPVALALAVALGLULEUPRO
36   PHETHRLEUMETHISPROLYSPROLYSGLU
37   GLUPROPROHISARGGLUVALPROGLUASN
38   GLUTHRPROVALASPTHRASNLEUILEGLU
39   LEUASPTHRASNASPASPASPALAALAALA
40   GLUASPPHEALAARGGLNARGLEULYSGLY
41   METLYSASPASPLYSGLUGLUGLUGLUASP
42   GLYTHRGLYSERPROGLNLEUASNASNARG

Entity 5, PIP2 - Formula weight is not available

1   2

Samples:

sample_1: enNTS1DM4, [13CH3-methionine], 66 uM; potassium phosphate 50 mM; NaCl 100 mM; DSS 20 uM; DDM 0.05%; NT8-13 500 uM; ML314 500 uM; Cysteine-free human beta-Arrestin 1 3A 151 uM; 08:0 PI(4,5)P2 130 uM

sample_conditions_1: ionic strength: 150 mM; pH: 7.4; pressure: 1 atm; temperature: 298 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-13C HMQCsample_1isotropicsample_conditions_1

Software:

SPARKY - chemical shift assignment, data analysis

TOPSPIN - data collection

qMDD - processing

NMR spectrometers:

  • Bruker AVANCE NEO 600 MHz