SASDDN6: Human stress-inducible heat shock 70 kDa protein 1, Hsp70 (Hspa1a...

Basic information

• Entry: Database: SASBDB / ID: SASDDN6

Sample

Human stress-inducible heat shock 70 kDa protein 1, Hsp70 (Hspa1a) T204A mutant dimer, in the presence of ATP

• Heat shock 70 kDa protein 1 (protein), HSPA1A_T204A, Homo sapiens

Function / homology

Function:

positive regulation of endoribonuclease activity / denatured protein binding / cellular heat acclimation / death receptor agonist activity / negative regulation of inclusion body assembly / Viral RNP Complexes in the Host Cell Nucleus / positive regulation of nucleotide-binding oligomerization domain containing 2 signaling pathway / C3HC4-type RING finger domain binding / positive regulation of microtubule nucleation / ATP-dependent protein disaggregase activity / misfolded protein binding / regulation of mitotic spindle assembly / negative regulation of mitochondrial outer membrane permeabilization involved in apoptotic signaling pathway / positive regulation of tumor necrosis factor-mediated signaling pathway / transcription regulator inhibitor activity / aggresome / lysosomal transport / cellular response to steroid hormone stimulus / mRNA catabolic process / regulation of protein ubiquitination / chaperone cofactor-dependent protein refolding / HSF1-dependent transactivation / response to unfolded protein / negative regulation of extrinsic apoptotic signaling pathway in absence of ligand / chaperone-mediated protein complex assembly / Regulation of HSF1-mediated heat shock response / Attenuation phase / cellular response to unfolded protein / negative regulation of endoplasmic reticulum stress-induced intrinsic apoptotic signaling pathway / ATP metabolic process / protein folding chaperone / inclusion body / negative regulation of protein ubiquitination / heat shock protein binding / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / centriole / positive regulation of RNA splicing / positive regulation of erythrocyte differentiation / AUF1 (hnRNP D0) binds and destabilizes mRNA / positive regulation of interleukin-8 production / G protein-coupled receptor binding / ATP-dependent protein folding chaperone / negative regulation of transforming growth factor beta receptor signaling pathway / PKR-mediated signaling / negative regulation of cell growth / histone deacetylase binding / transcription corepressor activity / disordered domain specific binding / unfolded protein binding / positive regulation of proteasomal ubiquitin-dependent protein catabolic process / virus receptor activity / positive regulation of NF-kappaB transcription factor activity / cellular response to heat / cellular response to oxidative stress / protein refolding / vesicle / ficolin-1-rich granule lumen / receptor ligand activity / protein stabilization / blood microparticle / nuclear speck / ribonucleoprotein complex / cadherin binding / negative regulation of cell population proliferation / focal adhesion / signaling receptor binding / centrosome / ubiquitin protein ligase binding / Neutrophil degranulation / positive regulation of gene expression / negative regulation of apoptotic process / perinuclear region of cytoplasm / enzyme binding / negative regulation of transcription by RNA polymerase II / endoplasmic reticulum / ATP hydrolysis activity / protein-containing complex / mitochondrion / RNA binding / extracellular space / extracellular exosome / extracellular region / nucleoplasm / ATP binding / nucleus / plasma membrane / cytoplasm / cytosol

Domain/homology:

Heat shock hsp70 proteins family signature 2. / Heat shock hsp70 proteins family signature 1. / Heat shock hsp70 proteins family signature 3. / Heat shock protein 70, conserved site / Heat shock protein 70kD, peptide-binding domain superfamily / Heat shock protein 70 family / Hsp70 protein / Heat shock protein 70kD, C-terminal domain superfamily / ATPase, nucleotide binding domain

Component:

Heat shock 70 kDa protein 1A

• Biological species: Homo sapiens (human)
• Citation: Journal: Mol Cell Proteomics / Year: 2019; Title: Human Stress-inducible Hsp70 Has a High Propensity to Form ATP-dependent Antiparallel Dimers That Are Differentially Regulated by Cochaperone Binding.; Authors: Filip Trcka / Michal Durech / Pavla Vankova / Josef Chmelik / Veronika Martinkova / Jiri Hausner / Alan Kadek / Julien Marcoux / Tomas Klumpler / Borivoj Vojtesek / Petr Muller / Petr Man / ; Abstract: Eukaryotic protein homeostasis (proteostasis) is largely dependent on the action of highly conserved Hsp70 molecular chaperones. Recent evidence indicates that, apart from conserved molecular allostery, Hsp70 proteins have retained and adapted the ability to assemble as functionally relevant ATP-bound dimers throughout evolution. Here, we have compared the ATP-dependent dimerization of DnaK, human stress-inducible Hsp70, Hsc70 and BiP Hsp70 proteins, showing that their dimerization propensities differ, with stress-inducible Hsp70 being predominantly dimeric in the presence of ATP. Structural analyses using hydrogen/deuterium exchange mass spectrometry, native electrospray ionization mass spectrometry and small-angle X-ray scattering revealed that stress-inducible Hsp70 assembles in solution as an antiparallel dimer with the intermolecular interface closely resembling the ATP-bound dimer interfaces captured in DnaK and BiP crystal structures. ATP-dependent dimerization of stress-inducible Hsp70 is necessary for its efficient interaction with Hsp40, as shown by experiments with dimerization-deficient mutants. Moreover, dimerization of ATP-bound Hsp70 is required for its participation in high molecular weight protein complexes detected , supporting its functional role As human cytosolic Hsp70 can interact with tetratricopeptide repeat (TPR) domain containing cochaperones, we tested the interaction of Hsp70 ATP-dependent dimers with Chip and Tomm34 cochaperones. Although Chip associates with intact Hsp70 dimers to form a larger complex, binding of Tomm34 disrupts the Hsp70 dimer and this event plays an important role in Hsp70 activity regulation. In summary, this study provides structural evidence of robust ATP-dependent antiparallel dimerization of human inducible Hsp70 protein and suggests a novel role of TPR domain cochaperones in multichaperone complexes involving Hsp70 ATP-bound dimers.

Contact author

• Tomas Klumpler (CEITEC - Central European Institute of Technology, Masaryk University)

Models

• Model #2029: ; Type: atomic / Chi-square value: 1.150 / P-value: 0.489949; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: Human stress-inducible heat shock 70 kDa protein 1, Hsp70 (Hspa1a) T204A mutant dimer, in the presence of ATP; Specimen concentration: 2.5 mg/ml
• Buffer: Name: 50mM HEPES, 150mM KCH3COO, 2mM MgCl2 / pH: 7.5

Entity #1080

Name: HSPA1A_T204A / Type: protein / Description: Heat shock 70 kDa protein 1 / Formula weight: 73.74 / Num. of mol.: 2 / Source: Homo sapiens / References: UniProt: P0DMV8
Sequence:

MSYYHHHHHH LESTSLYKKA GFGSENLYFQ SMAKAAAIGI DLGTTYSCVG VFQHGKVEII ANDQGNRTTP SYVAFTDTER LIGDAAKNQV ALNPQNTVFD AKRLIGRKFG DPVVQSDMKH WPFQVINDGD KPKVQVSYKG ETKAFYPEEI SSMVLTKMKE IAEAYLGYPV TNAVITVPAY FNDSQRQATK DAGVIAGLNV LRIINEPTAA AIAYGLDRTG KGERNVLIFD LGGGAFDVSI LTIDDGIFEV KATAGDTHLG GEDFDNRLVN HFVEEFKRKH KKDISQNKRA VRRLRTACER AKRTLSSSTQ ASLEIDSLFE GIDFYTSITR ARFEELCSDL FRSTLEPVEK ALRDAKLDKA QIHDLVLVGG STRIPKVQKL LQDFFNGRDL NKSINPDEAV AYGAAVQAAI LMGDKSENVQ DLLLLDVAPL SLGLETAGGV MTALIKRNST IPTKQTQIFT TYSDNQPGVL IQVYEGERAM TKDNNLLGRF ELSGIPPAPR GVPQIEVTFD IDANGILNVT ATDKSTGKAN KITITNDKGR LSKEEIERMV QEAEKYKAED EVQRERVSAK NALESYAFNM KSAVEDEGLK GKISEADKKK VLDKCQEVIS WLDANTLAEK DEFEHKRKEL EQVCNPIISG LYQGAGGPGP GGFGAQGPKG GSGSGPTIEE VD

Experimental information

• Beam: Instrument name: CEITEC Rigaku BioSAXS-1000 / City: Brno / 国: Czech Republic / Type of source: X-ray in house / Wavelength: 0.154 Å / Dist. spec. to detc.: 0.48 mm
• Detector: Name: Pilatus 100K / Pixsize x: 172 mm

Scan

Title: Human stress-inducible heat shock 70 kDa protein 1, Hsp70 (Hspa1a) T204A mutant dimer, in the presence of ATP
Measurement date: Feb 20, 2018 / Storage temperature: 4 °C / Cell temperature: 4 °C / Exposure time: 3600 sec. / Unit: 1/A /

	Min	Max
Q	0.0138	0.6314

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 130 /

	Min	Max
Q	0.0137969	0.199503
P(R) point	1	130
R	0	115.6

Result

Type of curve: single_conc /

	Experimental	Porod
MW	148 kDa	155 kDa
Volume	-	248 nm3

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	0.7457	0.0058	0.76	0.011
Radius of gyration, Rg	3.97 nm	0.2	4.01 nm	0.077

	Min	Max
D	-	11.56
Guinier point	2	14