BMRB Entry 19194

Chem Shift validation: AVS_full, LACS
BMRB Entry DOI: doi:10.13018/BMR19194

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Title:
Backbone resonance assignment of the alpha-subunit within the megadalton proteasome-activator complex
Deposition date:
2013-04-25
Original release date:
2013-08-13
Authors:
Mainz, Andi; Religa, Tomasz; Sprangers, Remco; Linser, Rasmus; Kay, Lewis; Reif, Bernd
Citation:

Citation: Mainz, Andi; Religa, Tomasz; Sprangers, Remco; Linser, Rasmus; Kay, Lewis; Reif, Bernd. "NMR spectroscopy of soluble protein complexes at one mega-dalton and beyond"  Angew. Chem. Int. Ed. 52, 8746-8751 (2013).
PubMed: 23873792

Assembly members:

Assembly members:
proteasome_alpha_subunit, polymer, 237 residues, 26115.8 Da.
proteasome_beta_subunit, polymer, 203 residues, 22271.8 Da.
proteasome_activator_PA26, polymer, 235 residues, 25229.7 Da.

Natural source:

Natural source:   Common Name: Thermoplasma acidophilum   Taxonomy ID: 2303   Superkingdom: Archaea   Kingdom: not available   Genus/species: Thermoplasma acidophilum

Experimental source:

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli   Vector: pETM-11

Entity Sequences (FASTA):

Entity Sequences (FASTA):
proteasome_alpha_subunit: GAMGMQQGQMAYDRAITVFS PDGRLFQVEYAREAVKKGST ALGMKFANGVLLISDKKVRS RLIEQNSIEKIQLIDDYVAA VTSGLVADARVLVDFARISA QQEKVTYGSLVNIENLVKRV ADQMQQYTQYGGVRPYGVSL IFAGIDQIGPRLFDCDPAGT INEYKATAIGSGKDAVVSFL EREYKENLPEKEAVTLGIKA LKSSLEEGEELKAPEIASIT VGNKYRIYDQEEVKKFL
proteasome_beta_subunit: TTTVGITLKDAVIMATERRV TMENFIMHKNGKKLFQIDTY TGMTIAGLVGDAQVLVRYMK AELELYRLQRRVNMPIEAVA TLLSNMLNQVKYMPYMVQLL VGGIDTAPHVFSIDAAGGSV EDIYASTGSGSPFVYGVLES QYSEKMTVDEGVDLVIRAIS AAKQRDSASGGMIDVAVITR KDGYVQLPTDQIESRIRKLG LIL
proteasome_activator_PA26: GAMGMPPKRAALIQNLRDSY TETSSFAVIEEWAAGTLQEI EGIAKAAAEAHGTIRNSTYG RAQAEKSPEQLLGVLQRYQD LCHNVYCQAETIRTVIAIRI PEHKEEDNLGVAVQHAVLKI IDELEIKTLGSGEKSGSGGA PTPIGMYALREYLSARSTVE DKLLGSVDAESGKTKGGSQS PSLLLELRQIDADFMLKVEL ATTHLSTMVRAVINAYLLNW KKLIQPRTGSDHMVS

Data sets:
Data typeCount
13C chemical shifts247
15N chemical shifts109
1H chemical shifts109

Additional metadata:

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

Entity Assembly IDEntity NameEntity ID
1alpha subunit1
2beta subunit2
311S activator3

Entities:

Entity 1, alpha subunit 237 residues - 26115.8 Da.

Original protein sequence. The first four residues in the entity sequence (GAMG: (-3)-(0)) are an overhang due to proteolytic tag removal.

1   GLYALAMETGLYMETGLNGLNGLYGLNMET
2   ALATYRASPARGALAILETHRVALPHESER
3   PROASPGLYARGLEUPHEGLNVALGLUTYR
4   ALAARGGLUALAVALLYSLYSGLYSERTHR
5   ALALEUGLYMETLYSPHEALAASNGLYVAL
6   LEULEUILESERASPLYSLYSVALARGSER
7   ARGLEUILEGLUGLNASNSERILEGLULYS
8   ILEGLNLEUILEASPASPTYRVALALAALA
9   VALTHRSERGLYLEUVALALAASPALAARG
10   VALLEUVALASPPHEALAARGILESERALA
11   GLNGLNGLULYSVALTHRTYRGLYSERLEU
12   VALASNILEGLUASNLEUVALLYSARGVAL
13   ALAASPGLNMETGLNGLNTYRTHRGLNTYR
14   GLYGLYVALARGPROTYRGLYVALSERLEU
15   ILEPHEALAGLYILEASPGLNILEGLYPRO
16   ARGLEUPHEASPCYSASPPROALAGLYTHR
17   ILEASNGLUTYRLYSALATHRALAILEGLY
18   SERGLYLYSASPALAVALVALSERPHELEU
19   GLUARGGLUTYRLYSGLUASNLEUPROGLU
20   LYSGLUALAVALTHRLEUGLYILELYSALA
21   LEULYSSERSERLEUGLUGLUGLYGLUGLU
22   LEULYSALAPROGLUILEALASERILETHR
23   VALGLYASNLYSTYRARGILETYRASPGLN
24   GLUGLUVALLYSLYSPHELEU

Entity 2, beta subunit 203 residues - 22271.8 Da.

1   THRTHRTHRVALGLYILETHRLEULYSASP
2   ALAVALILEMETALATHRGLUARGARGVAL
3   THRMETGLUASNPHEILEMETHISLYSASN
4   GLYLYSLYSLEUPHEGLNILEASPTHRTYR
5   THRGLYMETTHRILEALAGLYLEUVALGLY
6   ASPALAGLNVALLEUVALARGTYRMETLYS
7   ALAGLULEUGLULEUTYRARGLEUGLNARG
8   ARGVALASNMETPROILEGLUALAVALALA
9   THRLEULEUSERASNMETLEUASNGLNVAL
10   LYSTYRMETPROTYRMETVALGLNLEULEU
11   VALGLYGLYILEASPTHRALAPROHISVAL
12   PHESERILEASPALAALAGLYGLYSERVAL
13   GLUASPILETYRALASERTHRGLYSERGLY
14   SERPROPHEVALTYRGLYVALLEUGLUSER
15   GLNTYRSERGLULYSMETTHRVALASPGLU
16   GLYVALASPLEUVALILEARGALAILESER
17   ALAALALYSGLNARGASPSERALASERGLY
18   GLYMETILEASPVALALAVALILETHRARG
19   LYSASPGLYTYRVALGLNLEUPROTHRASP
20   GLNILEGLUSERARGILEARGLYSLEUGLY
21   LEUILELEU

Entity 3, 11S activator 235 residues - 25229.7 Da.

Orignal protein sequence. First four residues (GAMG: (-3)-(0)) are a protease cleavage artefact after tag removal.

1   GLYALAMETGLYMETPROPROLYSARGALA
2   ALALEUILEGLNASNLEUARGASPSERTYR
3   THRGLUTHRSERSERPHEALAVALILEGLU
4   GLUTRPALAALAGLYTHRLEUGLNGLUILE
5   GLUGLYILEALALYSALAALAALAGLUALA
6   HISGLYTHRILEARGASNSERTHRTYRGLY
7   ARGALAGLNALAGLULYSSERPROGLUGLN
8   LEULEUGLYVALLEUGLNARGTYRGLNASP
9   LEUCYSHISASNVALTYRCYSGLNALAGLU
10   THRILEARGTHRVALILEALAILEARGILE
11   PROGLUHISLYSGLUGLUASPASNLEUGLY
12   VALALAVALGLNHISALAVALLEULYSILE
13   ILEASPGLULEUGLUILELYSTHRLEUGLY
14   SERGLYGLULYSSERGLYSERGLYGLYALA
15   PROTHRPROILEGLYMETTYRALALEUARG
16   GLUTYRLEUSERALAARGSERTHRVALGLU
17   ASPLYSLEULEUGLYSERVALASPALAGLU
18   SERGLYLYSTHRLYSGLYGLYSERGLNSER
19   PROSERLEULEULEUGLULEUARGGLNILE
20   ASPALAASPPHEMETLEULYSVALGLULEU
21   ALATHRTHRHISLEUSERTHRMETVALARG
22   ALAVALILEASNALATYRLEULEUASNTRP
23   LYSLYSLEUILEGLNPROARGTHRGLYSER
24   ASPHISMETVALSER

Samples:

20S-11S_complex: proteasome alpha subunit, [U-13C; U-15N; U-2H], 3.0 mM; proteasome beta subunit, [U-2H], 3.0 mM; proteasome activator PA26, [U-2H], 4.5 mM; potassium phosphate 50 mM; sodium chloride 100 mM; DTT 1 mM; sodium azide 0.03 % w/v; glycerol, [U-2H], 30 % v/v; Cu(II)-EDTA 60 mM

sample_conditions_1: pH: 7.5; temperature: 303 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N HSQC via CP20S-11S_complexsedimentsample_conditions_1
2D 1H-15N HSQC via INEPT20S-11S_complexsedimentsample_conditions_1
3D hCOhNH via long-range CP20S-11S_complexsedimentsample_conditions_1
3D hCAhNH via long-range CP20S-11S_complexsedimentsample_conditions_1

Software:

TOPSPIN, Bruker Biospin - collection, data analysis, processing

SPARKY, Goddard - chemical shift assignment, data analysis

NMR spectrometers:

  • Bruker Avance 600 MHz

Related Database Links:

UniProtKB P25156 P28061 Q9U8G2
EMBL CAA42094.1 CAA42094 CAC12411 AAA72102.1 CAC11751 AAD50581.1 CBH15376
PDB
REF WP_010901695 NP_394085 WP_010901036 XP_822620
SP P25156 P28061
GB AAA72102 AAD50581 EAN77792
AlphaFold P25156 P28061

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