BMRB Entry 51927

Chem Shift validation: AVS_full
BMRB Entry DOI: doi:10.13018/BMR51927

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Title:
NT8-13 & ML314 bound enNTS1DM4 with G alpha iq & PIP2
Deposition date:
2023-04-18
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, 357 residues, 40611.26 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: GPGSGCTLSAEDKAAVERSK MIDRNLREDGEKAAREVKLL LLGAGESGKSTIVKQMKIIH EAGYSEEECKQYKAVVYSNT IQSIIAIIRAMGRLKIDFGD SARADDARQLFVLAGAAEEG FMTAELAGVIKRLWKDSGVQ ACFNRSREYQLNDSAAYYLN DLDRIAQPNYIPTQQDVLRT RVKTTGIVETHFTFKDLHFK MFDVGGQRSERKKWIHCFEG VTAIIFCVALSDYDLVLAED EEMNRMHESMKLFDSICNNK WFTDTSIILFLNKKDLFEEK IKKSPLTICYPEYAGSNTYE EAAAYIQCQFEDLNKRKDTK EIYTHFTCATDTKNVQFVFD AVTDVIIKNNLKEYNLV

Data sets:
Data typeCount
13C chemical shifts13
1H chemical shifts39

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
4enNTS1DM4, conformer D1
5enNTS1DM4, conformer E1
6NT8-132
7ML3143
8Gaiq4
9PIP25

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, Gaiq 357 residues - 40611.26 Da.

G alpha iq is a chimera of G alpha i1 with the six C-terminal residues from G alpha q (KEYNLV)

1   GLYPROGLYSERGLYCYSTHRLEUSERALA
2   GLUASPLYSALAALAVALGLUARGSERLYS
3   METILEASPARGASNLEUARGGLUASPGLY
4   GLULYSALAALAARGGLUVALLYSLEULEU
5   LEULEUGLYALAGLYGLUSERGLYLYSSER
6   THRILEVALLYSGLNMETLYSILEILEHIS
7   GLUALAGLYTYRSERGLUGLUGLUCYSLYS
8   GLNTYRLYSALAVALVALTYRSERASNTHR
9   ILEGLNSERILEILEALAILEILEARGALA
10   METGLYARGLEULYSILEASPPHEGLYASP
11   SERALAARGALAASPASPALAARGGLNLEU
12   PHEVALLEUALAGLYALAALAGLUGLUGLY
13   PHEMETTHRALAGLULEUALAGLYVALILE
14   LYSARGLEUTRPLYSASPSERGLYVALGLN
15   ALACYSPHEASNARGSERARGGLUTYRGLN
16   LEUASNASPSERALAALATYRTYRLEUASN
17   ASPLEUASPARGILEALAGLNPROASNTYR
18   ILEPROTHRGLNGLNASPVALLEUARGTHR
19   ARGVALLYSTHRTHRGLYILEVALGLUTHR
20   HISPHETHRPHELYSASPLEUHISPHELYS
21   METPHEASPVALGLYGLYGLNARGSERGLU
22   ARGLYSLYSTRPILEHISCYSPHEGLUGLY
23   VALTHRALAILEILEPHECYSVALALALEU
24   SERASPTYRASPLEUVALLEUALAGLUASP
25   GLUGLUMETASNARGMETHISGLUSERMET
26   LYSLEUPHEASPSERILECYSASNASNLYS
27   TRPPHETHRASPTHRSERILEILELEUPHE
28   LEUASNLYSLYSASPLEUPHEGLUGLULYS
29   ILELYSLYSSERPROLEUTHRILECYSTYR
30   PROGLUTYRALAGLYSERASNTHRTYRGLU
31   GLUALAALAALATYRILEGLNCYSGLNPHE
32   GLUASPLEUASNLYSARGLYSASPTHRLYS
33   GLUILETYRTHRHISPHETHRCYSALATHR
34   ASPTHRLYSASNVALGLNPHEVALPHEASP
35   ALAVALTHRASPVALILEILELYSASNASN
36   LEULYSGLUTYRASNLEUVAL

Entity 5, PIP2 - Formula weight is not available

1   2

Samples:

sample_1: enNTS1DM4, [13CH3-methionine], 64 uM; potassium phosphate 50 mM; NaCl 100 mM; DSS 20 uM; DDM 0.05%; NT8-13 500 uM; ML314 500 uM; G alpha i/q chimera 145 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