BMRB Entry 50243

Title:
Backbone (1H, 13C and 15N) Chemical Shift Assignments and 15N Relaxation Parameters for protein kinase Inhibitor alpha (PKIa) bound to cAMP-dependent protein kinase A   PubMed: 32338601
Deposition date:
2020-04-23
Original release date:
2020-05-15
Authors:
Olivieri, Cristina; Li, Geoffrey; Veliparambil Subrahmanian, Manu; Veglia, Gianluigi
Citation:

Citation: Olivieri, Cristina; Wang, Yingjie; Li, Geoffrey; Veliparambil Subrahmanian, Manu; Kim, Jonggul; Stultz, Benjamin; Neibergall, Matthew; Porcelli, Fernando; Muretta, Joseph; Thomas, David; Gao, Jiali; Blumenthal, Donald; Taylor, Susan; Veglia, Gianluigi. "Multi-state recognition pathway of the intrinsically disordered protein kinase inhibitor by protein kinase A"  eLIFE 9, e55607-e55607 (2020).

Assembly members:

Assembly members:
entity_1, polymer, 75 residues, 7857.33 Da.
entity_2, polymer, 350 residues, 40439.43 Da.
entity_ANP, non-polymer, 506.196 Da.
entity_MG, non-polymer, 24.305 Da.

Natural source:

Natural source:   Common Name: rabbit   Taxonomy ID: 9986   Superkingdom: Eukaryota   Kingdom: Metazoa   Genus/species: Oryctolagus cuniculus

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pT7-7

Experimental source:

Natural source:   Common Name: rabbit   Taxonomy ID: 9986   Superkingdom: Eukaryota   Kingdom: Metazoa   Genus/species: Oryctolagus cuniculus

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pT7-7

Data sets:
Data typeCount
13C chemical shifts201
15N chemical shifts74
1H chemical shifts222
T1 relaxation values69
T2 relaxation values71
heteronuclear NOE values60

Additional metadata:

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

Assembly:

Entity Assembly IDEntity NameEntity ID
1PKIA1
2PKA-C2
3AMPPNP3
4Mg2+4

Entities:

Entity 1, PKIA 75 residues - 7857.33 Da.

1   THRASPVALGLUTHRTHRTYRALAASPPHE
2   ILEALASERGLYARGTHRGLYARGARGASN
3   ALAILEHISASPILELEUVALSERSERALA
4   SERGLYASNSERASNGLULEUALALEULYS
5   LEUALAGLYLEUASPILEASNLYSTHRGLU
6   GLYGLUGLUASPALAGLNARGSERSERTHR
7   GLUGLNSERGLYGLUALAGLNGLYGLUALA
8   ALALYSSERGLUSER

Entity 2, PKA-C 350 residues - 40439.43 Da.

1   GLYASNALAALAALAALALYSLYSGLYSER
2   GLUGLNGLUSERVALLYSGLUPHELEUALA
3   LYSALALYSGLUASPPHELEULYSLYSTRP
4   GLUTHRPROSERGLNASNTHRALAGLNLEU
5   ASPGLNPHEASPARGILELYSTHRLEUGLY
6   THRGLYSERPHEGLYARGVALMETLEUVAL
7   LYSHISLYSGLUSERGLYASNHISTYRALA
8   METLYSILELEUASPLYSGLNLYSVALVAL
9   LYSLEULYSGLNILEGLUHISTHRLEUASN
10   GLULYSARGILELEUGLNALAVALASNPHE
11   PROPHELEUVALLYSLEUGLUPHESERPHE
12   LYSASPASNSERASNLEUTYRMETVALMET
13   GLUTYRVALALAGLYGLYGLUMETPHESER
14   HISLEUARGARGILEGLYARGPHESERGLU
15   PROHISALAARGPHETYRALAALAGLNILE
16   VALLEUTHRPHEGLUTYRLEUHISSERLEU
17   ASPLEUILETYRARGASPLEULYSPROGLU
18   ASNLEULEUILEASPGLNGLNGLYTYRILE
19   GLNVALTHRASPPHEGLYPHEALALYSARG
20   VALLYSGLYARGTHRTRPTHRLEUCYSGLY
21   THRPROGLUTYRLEUALAPROGLUILEILE
22   LEUSERLYSGLYTYRASNLYSALAVALASP
23   TRPTRPALALEUGLYVALLEUILETYRGLU
24   METALAALAGLYTYRPROPROPHEPHEALA
25   ASPGLNPROILEGLNILETYRGLULYSILE
26   VALSERGLYLYSVALARGPHEPROSERHIS
27   PHESERSERASPLEULYSASPLEULEUARG
28   ASNLEULEUGLNVALASPLEUTHRLYSARG
29   PHEGLYASNLEULYSASNGLYVALASNASP
30   ILELYSASNHISLYSTRPPHEALATHRTHR
31   ASPTRPILEALAILETYRGLNARGLYSVAL
32   GLUALAPROPHEILEPROLYSPHELYSGLY
33   PROGLYASPTHRSERASNPHEASPASPTYR
34   GLUGLUGLUGLUILEARGVALSERILEASN
35   GLULYSCYSGLYLYSGLUPHETHRGLUPHE

Entity 3, AMPPNP - C10 H17 N6 O12 P3 - 506.196 Da.

1   ANP

Entity 4, Mg2+ - Mg - 24.305 Da.

1   MG

Samples:

sample_1: cAMP-dependent protein kinase A Inhibitor alpha (PKIA), [U-13C; U-15N; U-2H], 0.225 mM; potassium phosphate 20 mM; potassium chloride 90 mM; magnesium chloride 10 mM; DTT 10 mM; sodium azide 1 mM; cAMP-dependent protein kinase A catalytic subunit alpha (PKA-C), [U-2H], 0.225 mM; AMPPNP 12 mM

sample_2: cAMP-dependent protein kinase A Inhibitor alpha (PKIA), [U-15N; U-2H], 0.225 mM; potassium phosphate 20 mM; potassium chloride 90 mM; magnesium chloride 10 mM; DTT 10 mM; sodium azide 1 mM; cAMP-dependent protein kinase A catalytic subunit alpha (PKA-C), [U-2H], 0.15-0.3 mM; AMPPNP 12 mM

sample_conditions_1: ionic strength: 0.24 M; pH: 6.5; pressure: 1 atm; temperature: 300 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N TROSY HSQCsample_1isotropicsample_conditions_1
3D HNCAsample_1isotropicsample_conditions_1
3D HNCACBsample_1isotropicsample_conditions_1
3D HN(CO)CAsample_1isotropicsample_conditions_1
3D 1H-15N NOESY-HSQCsample_1isotropicsample_conditions_1
T1/R1 relaxationsample_2isotropicsample_conditions_1
T2/R2 relaxationsample_2isotropicsample_conditions_1
1H-15N heteronoesample_2isotropicsample_conditions_1

Software:

TOPSPIN - collection

NMRPipe - data analysis, processing

NMRDraw - processing

SPARKY vNMRFam - chemical shift assignment, chemical shift calculation, data analysis, peak picking

NMR spectrometers:

  • Bruker Avance 700 MHz
  • Bruker Avance III 850 MHz
  • Bruker Avance III 900 MHz
  • Varian INOVA 600 MHz

Related Database Links:

UNP P61926 P05132
AlphaFold P04541 Q9JID0

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