Reversible thermal transition in GrpE, the nucleotide exchange factor of the DnaK heat-shock system.


Abstract

DnaK, a Hsp70 acting in concert with its co-chaperones DnaJ and GrpE, is essential for Escherichia coli to survive environmental stress, including exposure to elevated temperatures. Here we explored the influence of temperature on the structure of the individual components and the functional properties of the chaperone system. GrpE undergoes extensive but fully reversible conformational changes in the physiologically relevant temperature range (transition midpoint at approximately 48 degrees C), as observed with both circular dichroism measurements and differential scanning calorimetry, whereas no thermal transitions occur in DnaK and DnaJ between 15 degrees C and 48 degrees C. The conformational changes in GrpE appear to be important in controlling the interconversion of T-state DnaK (ATP-liganded, low affinity for polypeptide substrates) and R-state DnaK (ADP-liganded, high affinity for polypeptide substrates). The rate of the T --> R conversion of DnaK due to DnaJ-triggered ATP hydrolysis follows an Arrhenius temperature dependence. In contrast, the rate of the R --> T conversion due to GrpE-catalyzed ADP/ATP exchange increases progressively less with increasing temperature and even decreases at temperatures above approximately 40 degrees C, indicating a temperature-dependent reversible inactivation of GrpE. At heat-shock temperatures, the reversible structural changes of GrpE thus shift DnaK toward its high-affinity R state. Study holds ProTherm entries: 10527, 10528 Extra Details: environmental stress; conformational changes; affinity;,temperature dependence

Submission Details

ID: feXxGnES3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:40 p.m.

Version: 1

Publication Details
Grimshaw JP;Jelesarov I;Schönfeld HJ;Christen P,J. Biol. Chem. (2001) Reversible thermal transition in GrpE, the nucleotide exchange factor of the DnaK heat-shock system. PMID:11084044
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1DG4 1999-12-08 NMR STRUCTURE OF THE SUBSTRATE BINDING DOMAIN OF DNAK IN THE APO FORM
2KHO 2009-05-12 NMR-RDC / XRAY structure of E. coli HSP70 (DNAK) chaperone (1-605) complexed with ADP and substrate
1Q5L 2003-11-04 NMR structure of the substrate binding domain of DnaK bound to the peptide NRLLLTG
2BPR 1999-03-02 NMR STRUCTURE OF THE SUBSTRATE BINDING DOMAIN OF DNAK, 25 STRUCTURES
1BPR 1999-03-02 NMR STRUCTURE OF THE SUBSTRATE BINDING DOMAIN OF DNAK, MINIMIZED AVERAGE STRUCTURE
4F01 2013-04-17 1.4 Crystal structure of an artificial dimeric DnaK complex
4HY9 2013-04-17 1.55 Crystal structure of the substrate binding domain of E.coli DnaK in complex with pyrrhocoricin_LYZZ (residues 1 to 11)
4JNF 2013-05-29 1.62 Allosteric opening of the polypeptide-binding site when an Hsp70 binds ATP
4EZP 2013-04-17 1.65 Crystal structure of the substrate binding domain of E.coli DnaK in complex with A3-APO(residues 1 to 20)
4EZX 2013-04-17 1.7 Crystal structure of the substrate binding domain of E.coli DnaK in complex with the designer peptide NRLMLTG
4HYB 2013-04-17 1.7 Crystal structure of the substrate binding domain of E.coli DnaK in complex with pyrrhocoricin_LYZI (residues 1 to 10)
4R5K 2014-09-10 1.75 Crystal structure of the DnaK C-terminus (Dnak-SBD-B)
4JWC 2013-11-13 1.8 Crystal structure of the substrate binding domain of E.coli DnaK in complex with bovine Bac7(1-16)
4EZN 2013-04-17 1.8 Crystal structure of the substrate binding domain of E.coli DnaK in complex with pyrrhocoricin
4EZW 2013-04-17 1.8 Crystal structure of the substrate binding domain of E.coli DnaK in complex with the designer peptide NRLLLTG
4EZY 2013-04-17 1.85 Crystal structure of the substrate binding domain of E.coli DnaK in complex with the designer peptide NRLILTG
4EZO 2013-04-17 1.9 Crystal structure of the substrate binding domain of E.coli DnaK in complex with PR-39 (residues 1 to 15)
4EZR 2013-04-17 1.9 Crystal structure of the substrate binding domain of E.coli DnaK in complex with the C-terminal part of drosocin (residues 12 to 19)
4EZU 2013-04-24 1.9 Crystal structure of the substrate binding domain of E.coli DnaK in complex with PR-bombesin in space group I222
4JWI 2013-11-13 1.9 Crystal structure of the substrate binding domain of E.coli DnaK in complex with sheep Bac7(35-43)
4EZS 2013-04-24 1.9 Crystal structure of the substrate binding domain of E.coli DnaK in complex with metchnikowin (residues 20 to 26)
4E81 2012-05-30 1.9 Crystal structure of the substrate binding domain of E.coli DnaK in complex with a short apidaecin peptide
4JWE 2013-11-13 1.95 Crystal structure of the substrate binding domain of E.coli DnaK in complex with sheep Bac7(1-21)
4F00 2013-04-17 1.95 Crystal structure of the substrate binding domain of E.coli DnaK in complex with an apidaecin fragment from the bumblebee (residues 3 to 11)
4JWD 2013-11-13 1.95 Crystal structure of the substrate binding domain of E.coli DnaK in complex with bovine Bac7(15-28)
4JNE 2013-05-29 1.96 Allosteric opening of the polypeptide-binding site when an Hsp70 binds ATP
4R5I 2014-09-10 1.97 Crystal structure of the DnaK C-terminus with the substrate peptide NRLLLTG
4EZT 2013-04-17 2.0 Crystal structure of the substrate binding domain of E.coli DnaK in complex with heliocin (residues 14 to 21)
1DKZ 1996-12-07 2.0 THE SUBSTRATE BINDING DOMAIN OF DNAK IN COMPLEX WITH A SUBSTRATE PEPTIDE, DETERMINED FROM TYPE 1 NATIVE CRYSTALS
1DKX 1996-12-07 2.0 THE SUBSTRATE BINDING DOMAIN OF DNAK IN COMPLEX WITH A SUBSTRATE PEPTIDE, DETERMINED FROM TYPE 1 SELENOMETHIONYL CRYSTALS
4EZQ 2013-04-17 2.0 Crystal structure of the substrate binding domain of E.coli DnaK in complex with the C-terminal part of pyrrhocoricin (residues 12 to 20)
4EZZ 2013-04-17 2.05 Crystal structure of the substrate binding domain of E.coli DnaK in complex with the designer peptide ELPLVKI
4EZV 2013-04-24 2.1 Crystal structure of the substrate binding domain of E.coli DnaK in complex with PR-bombesin in space group P21212
3DPO 2009-03-10 2.1 Crystal structure of the substrate binding domain of E. coli DnaK in complex with a short pyrrhocoricin-derived inhibitor peptide
3QNJ 2011-03-23 2.28 Crystal structure of the substrate binding domain of E.coli DnaK in complex with the antimicrobial peptide oncocin
4JN4 2013-05-29 2.3 Allosteric opening of the polypeptide-binding site when an Hsp70 binds ATP
4R5J 2014-09-10 2.36 Crystal structure of the DnaK C-terminus (Dnak-SBD-A)
4B9Q 2012-11-14 2.4 Open conformation of ATP-bound Hsp70 homolog DnaK
3DPP 2009-03-10 2.5 Crystal structure of the substrate binding domain of E. coli DnaK in complex with a long pyrrhocoricin-derived inhibitor peptide (form A)
3DPQ 2009-03-10 2.6 Crystal structure of the substrate binding domain of E. coli DnaK in complex with a long pyrrhocoricin-derived inhibitor peptide (form B)
1DKG 1997-08-20 2.8 CRYSTAL STRUCTURE OF THE NUCLEOTIDE EXCHANGE FACTOR GRPE BOUND TO THE ATPASE DOMAIN OF THE MOLECULAR CHAPERONE DNAK
1DKY 1996-12-07 2.8 THE SUBSTRATE BINDING DOMAIN OF DNAK IN COMPLEX WITH A SUBSTRATE PEPTIDE, DETERMINED FROM TYPE 2 NATIVE CRYSTALS
5OOW 2018-04-25 2.9 Crystal structure of lobe II from the nucleotide binding domain of DnaK in complex with AMPPCP
4R5L 2014-09-10 2.97 Crystal structure of the DnaK C-terminus (Dnak-SBD-C)
5NRO 2018-01-17 3.25 Structure of full-length DnaK with bound J-domain
4R5G 2014-09-10 3.45 Crystal structure of the DnaK C-terminus with the inhibitor PET-16

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.3 Protein GrpE A4WDH8 GRPE_ENT38
182.6 A,B Protein GrpE B5RD90 GRPE_SALG2
183.6 A,B Protein GrpE Q7CPZ4 GRPE_SALTY
183.6 A,B Protein GrpE Q8XEY8 GRPE_SALTI
183.6 A,B Protein GrpE B4TS61 GRPE_SALSV
183.6 A,B Protein GrpE B5BE99 GRPE_SALPK
183.6 A,B Protein GrpE Q5PFG9 GRPE_SALPA
183.6 A,B Protein GrpE B4T2B9 GRPE_SALNS
183.6 A,B Protein GrpE B4TE57 GRPE_SALHS
183.6 A,B Protein GrpE B5QUG9 GRPE_SALEP
197.0 A,B Protein GrpE B1LPC1 GRPE_ECOSM
197.0 A,B Protein GrpE Q0T181 GRPE_SHIF8
197.0 A,B Protein GrpE B5Z231 GRPE_ECO5E
198.0 A,B Protein GrpE Q1R8B1 GRPE_ECOUT
198.0 A,B Protein GrpE B7MYA6 GRPE_ECO81
198.0 A,B Protein GrpE B7MIV1 GRPE_ECO45
198.0 A,B Protein GrpE A7ZQ54 GRPE_ECO24
198.0 A,B Protein GrpE Q3YYM5 GRPE_SHISS
198.0 A,B Protein GrpE Q7C0D0 GRPE_SHIFL
198.0 A,B Protein GrpE Q32CX5 GRPE_SHIDS
198.0 A,B Protein GrpE Q31XD2 GRPE_SHIBS
198.0 A,B Protein GrpE B2TYN5 GRPE_SHIB3
198.0 A,B Protein GrpE B6I635 GRPE_ECOSE
198.0 A,B Protein GrpE B7N6J9 GRPE_ECOLU
198.0 A,B Protein GrpE B1IVM0 GRPE_ECOLC
198.0 A,B Protein GrpE Q8FEY9 GRPE_ECOL6
198.0 A,B Protein GrpE Q0TEM6 GRPE_ECOL5
198.0 A,B Protein GrpE A8A3C0 GRPE_ECOHS
198.0 A,B Protein GrpE B7M983 GRPE_ECO8A
198.0 A,B Protein GrpE B7NSB2 GRPE_ECO7I
198.0 A,B Protein GrpE Q7ABI1 GRPE_ECO57
198.0 A,B Protein GrpE B7LDK2 GRPE_ECO55
198.0 A,B Protein GrpE B7UH62 GRPE_ECO27
199.0 A,B Protein GrpE P09372 GRPE_ECOLI
199.0 A,B Protein GrpE B1XBT4 GRPE_ECODH
199.0 A,B Protein GrpE C4ZYN1 GRPE_ECOBW
90.6 D Protein GrpE C3LTA5 DNAK_VIBCM
90.6 D Protein GrpE O34241 DNAK_VIBCH
90.6 D Protein GrpE A5F361 DNAK_VIBC3
90.6 D Protein GrpE P57870 DNAK_PASMU
92.4 D Protein GrpE C5B7L7 DNAK_EDWI9
91.9 D Protein GrpE Q6D0B7 DNAK_PECAS
92.2 D Protein GrpE C6DF10 DNAK_PECCP
92.1 D Protein GrpE Q2NVZ1 DNAK_SODGM
94.0 D Protein GrpE A8G9K8 DNAK_SERP5
93.7 D Protein GrpE B2VGR9 DNAK_ERWT9
93.7 D Protein GrpE Q7N8Y4 DNAK_PHOLL
95.0 D Protein GrpE A1JJD5 DNAK_YERE8
95.3 D Protein GrpE B1JL04 DNAK_YERPY
95.3 D Protein GrpE Q66ET0 DNAK_YERPS
95.3 D Protein GrpE A4TQF9 DNAK_YERPP
95.3 D Protein GrpE Q1CMV7 DNAK_YERPN
95.3 D Protein GrpE A9R015 DNAK_YERPG
95.3 D Protein GrpE Q8ZIM7 DNAK_YERPE
95.3 D Protein GrpE B2K3M0 DNAK_YERPB
95.3 D Protein GrpE Q1C0J9 DNAK_YERPA
95.3 D Protein GrpE A7FME3 DNAK_YERP3
96.6 D Protein GrpE A9MR77 DNAK_SALAR
96.3 D Protein GrpE A4W6D5 DNAK_ENT38
96.6 D Protein GrpE A6T4F4 DNAK_KLEP7
96.6 D Protein GrpE B5Y242 DNAK_KLEP3
96.9 D Protein GrpE Q56073 DNAK_SALTY
96.9 D Protein GrpE Q8Z9R1 DNAK_SALTI
96.9 D Protein GrpE B4TVZ5 DNAK_SALSV
96.9 D Protein GrpE B5BLH8 DNAK_SALPK
96.9 D Protein GrpE C0Q4F3 DNAK_SALPC
96.9 D Protein GrpE A9MXI2 DNAK_SALPB
96.9 D Protein GrpE Q5PDJ5 DNAK_SALPA
96.9 D Protein GrpE B4T6D6 DNAK_SALNS
96.9 D Protein GrpE B4TIB4 DNAK_SALHS
96.9 D Protein GrpE B5RF08 DNAK_SALG2
96.9 D Protein GrpE B5R5I2 DNAK_SALEP
96.9 D Protein GrpE B5FHA6 DNAK_SALDC
96.9 D Protein GrpE Q57TP3 DNAK_SALCH
96.9 D Protein GrpE B5F6Y8 DNAK_SALA4
97.4 D Protein GrpE A8ALU3 DNAK_CITK8
97.7 D Protein GrpE A7MIK5 DNAK_CROS8
99.5 D Protein GrpE Q83MH5 DNAK_SHIFL
99.5 D Protein GrpE Q0T8H6 DNAK_SHIF8
99.7 D Protein GrpE Q3Z601 DNAK_SHISS
99.7 D Protein GrpE Q32KA5 DNAK_SHIDS
99.7 D Protein GrpE Q326K7 DNAK_SHIBS
99.7 D Protein GrpE Q1RGI8 DNAK_ECOUT
99.7 D Protein GrpE P0A6Y8 DNAK_ECOLI
99.7 D Protein GrpE B1IRG0 DNAK_ECOLC
99.7 D Protein GrpE P0A6Y9 DNAK_ECOL6
99.7 D Protein GrpE Q0TLX5 DNAK_ECOL5
99.7 D Protein GrpE A1A766 DNAK_ECOK1
99.7 D Protein GrpE A7ZVV7 DNAK_ECOHS
99.7 D Protein GrpE P0A6Z0 DNAK_ECO57
99.7 D Protein GrpE A7ZHA4 DNAK_ECO24