brown fat cell proliferation / protein localization to perinuclear region of cytoplasm / regulation of Ras protein signal transduction / protein targeting to vacuole involved in autophagy / response to mitochondrial depolarisation / aggrephagy / Lewy body / negative regulation of toll-like receptor 4 signaling pathway / amphisome / regulation of protein complex stability ...brown fat cell proliferation / protein localization to perinuclear region of cytoplasm / regulation of Ras protein signal transduction / protein targeting to vacuole involved in autophagy / response to mitochondrial depolarisation / aggrephagy / Lewy body / negative regulation of toll-like receptor 4 signaling pathway / amphisome / regulation of protein complex stability / endosome organization / autophagy of mitochondrion / pexophagy / membraneless organelle assembly / phagophore assembly site / ubiquitin-modified protein reader activity / regulation of mitochondrion organization / regulation of canonical NF-kappaB signal transduction / Nuclear events mediated by NFE2L2 / endosomal transport / aggresome / intracellular membraneless organelle / K63-linked polyubiquitin modification-dependent protein binding / negative regulation of ferroptosis / temperature homeostasis / cellular response to stress / autolysosome / molecular sequestering activity / immune system process / mitophagy / energy homeostasis / sperm midpiece / inclusion body / signaling adaptor activity / negative regulation of protein ubiquitination / ionotropic glutamate receptor binding / positive regulation of autophagy / p75NTR recruits signalling complexes / SH2 domain binding / NF-kB is activated and signals survival / autophagosome / protein sequestering activity / Pexophagy / NRIF signals cell death from the nucleus / protein kinase C binding / sarcomere / positive regulation of long-term synaptic potentiation / ubiquitin binding / PINK1-PRKN Mediated Mitophagy / response to ischemia / positive regulation of protein localization to plasma membrane / macroautophagy / P-body / protein catabolic process / molecular condensate scaffold activity / PML body / receptor tyrosine kinase binding / autophagy / Interleukin-1 signaling / protein import into nucleus / KEAP1-NFE2L2 pathway / Signaling by ALK fusions and activated point mutants / late endosome / intracellular protein localization / signaling receptor activity / Neddylation / ubiquitin-dependent protein catabolic process / protein-macromolecule adaptor activity / transcription by RNA polymerase II / cell differentiation / intracellular signal transduction / positive regulation of apoptotic process / intracellular membrane-bounded organelle / apoptotic process / ubiquitin protein ligase binding / protein-containing complex binding / protein kinase binding / glutamatergic synapse / enzyme binding / endoplasmic reticulum / negative regulation of transcription by RNA polymerase II / positive regulation of transcription by RNA polymerase II / mitochondrion / extracellular exosome / zinc ion binding / nucleoplasm / identical protein binding / cytosol / cytoplasm Similarity search - Function
Sequestosome-1 / Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa / Ubiquitin-binding protein p62 ...Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa / Ubiquitin-binding protein p62 / EBI3-associated protein of 60 kDa / p60 / EBIAP
Mass: 5744.406 Da / Num. of mol.: 1 / Fragment: UBA domain (UNP residues 387-436) Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SQSTM1, ORCA, OSIL / Plasmid: pGEX-4T-1 / Species (production host): Escherichia coli / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / References: UniProt: Q13501
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Experimental details
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Experiment
Experiment
Method: SOLUTION NMR
NMR experiment
Conditions-ID
Experiment-ID
Solution-ID
Type
1
1
1
2D 1H-15N HSQC
1
2
1
3D 1H-15N NOESY
1
3
2
2D 1H-13C HSQC
1
4
2
3DCBCA(CO)NH
1
5
2
3D HN(CA)CB
1
6
2
3D HNCO
1
7
2
3DHN(CA)CO
1
8
2
3D (H)CCH-TOCSY
1
9
2
3D 1H-13C NOESY
1
10
1
3D 1H-15N TOCSY
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Sample preparation
Details
Solution-ID
Contents
Solvent system
1
1 mM [U-100% 15N] Sequestosome-1, 50 mM potassium phosphate, 50 mM sodium chloride, 90% H2O/10% D2O
90% H2O/10% D2O
2
1 mM [U-100% 13C; U-100% 15N] Sequestosome-1, 50 mM potassium phosphate, 50 mM sodium chloride, 90% H2O/10% D2O
90% H2O/10% D2O
Sample
Conc. (mg/ml)
Component
Isotopic labeling
Solution-ID
1mM
Sequestosome-1
[U-100% 15N]
1
50mM
potassiumphosphate
1
50mM
sodiumchloride
1
1mM
Sequestosome-1
[U-100% 13C; U-100% 15N]
2
50mM
potassiumphosphate
2
50mM
sodiumchloride
2
Sample conditions
pH: 7 / Pressure: ambient atm / Temperature: 298 K
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NMR measurement
NMR spectrometer
Type: Bruker Avance / Manufacturer: Bruker / Model: AVANCE / Field strength: 600 MHz
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Processing
NMR software
Name
Version
Developer
Classification
X-PLOR NIH
2.14
Schwieters, Kuszewski, TjandraandClore
structuresolution
TALOS
Cornilescu, DelaglioandBax
dataanalysis
XwinNMR
BrukerBiospin
collection
XwinNMR
BrukerBiospin
processing
CcpNMR
1.0.10
CCPN
chemicalshiftassignment
CcpNMR
1.0.10
CCPN
dataanalysis
CcpNMR
1.0.10
CCPN
peakpicking
X-PLOR NIH
2.14
Schwieters, Kuszewski, TjandraandClore
refinement
Refinement
Method: simulated annealing / Software ordinal: 1
NMR constraints
NOE constraints total: 1078 / NOE intraresidue total count: 360 / NOE long range total count: 213 / NOE medium range total count: 246 / NOE sequential total count: 259 / Protein phi angle constraints total count: 36 / Protein psi angle constraints total count: 36
NMR representative
Selection criteria: closest to the average
NMR ensemble
Conformer selection criteria: structures with the lowest energy Conformers calculated total number: 100 / Conformers submitted total number: 30
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