PXD044739

PXD044739 is an original dataset announced via ProteomeXchange.

Title	Analysis of reconstituted tripartite complex supports avidity-based recruitment of Hsp70 by substrate bound J-domain protein
Description	Hsp70 are ubiquitous, versatile molecular chaperones that cyclically interact with substrate protein(s). The initial step requires synergistic interaction of a substrate and a J-domain protein (JDP) cochaperone, via its J-domain, with Hsp70 to stimulate hydrolysis of its bound ATP. This hydrolysis drives conformational changes in Hsp70 that stabilize substrate binding. However, because of the transient nature of substrate and JDP interactions, this key step is not well understood. Here we leverage a well characterized Hsp70 system specialized for iron-sulfur cluster biogenesis, which like many systems, has a JDP that binds substrate on its own. Utilizing an ATPase-deficient Hsp70 variant, we isolated a Hsp70- JDP-substrate tripartite complex. Complex formation and stability depended on residues previously identified as essential for bipartite interactions: JDP-substrate, Hsp70-substrate and J-domain-Hsp70. Computational docking based on the established J-domain-Hsp70(ATP) interaction placed the substrate close to its predicted position in the peptide-binding cleft, with the JDP having the same architecture as when in a bipartite complex with substrate. Together, our results indicate that the structurally rigid JDP- substrate complex recruits Hsp70(ATP) via precise positioning of J-domain and substrate at their respective interaction sites - resulting in functionally high affinity (i.e., avidity). The exceptionally high avidity observed for this specialized system may be unusual because of the rigid architecture of its JDP and the additional JDP-Hsp70 interaction site uncovered in this study. However, functionally important avidity driven by JDP-substrate interactions is likely sufficient to explain synergistic ATPase stimulation and efficient substrate trapping in many Hsp70 systems.
HostingRepository	PRIDE
AnnounceDate	2024-02-19
AnnouncementXML	Submission_2024-02-18_18:27:36.733.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Igor Grochowina
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	TripleTOF 5600; Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2023-08-22 07:19:27	ID requested
⏵ 1	2024-02-18 18:27:37	announced

10.1016/J.JMB.2023.168283;

submitter keyword: Hsc20, Hsp70 ATPase cycle, substrate delivery, Ssq1,molecular chaperones

Jaroslaw Marszalek
contact affiliation	Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, 58 Abrahama St., 80-307 Gdansk, Poland
contact email	jaroslaw.marszalek@ug.edu.pl
lab head
Igor Grochowina
contact affiliation	Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, 58 Abrahama St., 80-307 Gdansk, Poland
contact email	igor.grochowina@phdstud.ug.edu.pl
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD044739
     1. Label: PRIDE project
     2. Name: Analysis of reconstituted tripartite complex supports avidity-based recruitment of Hsp70 by substrate bound J-domain protein