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Disulfide mapping of J-chain by crosslinking MS

ABSTRACT: Here we show how J-chain (JC) outcompetes the sixth IgM subunit during assembly. We looked at the disulfide re-arrangements in JC and showed that it exists as an unstructured protein with non-native disulfides before its insertion into IgM.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Yevheniia Bushman

LAB HEAD: Johannes Buchner

PROVIDER: PXD048614 | Pride | 2024-12-04

REPOSITORIES: pride

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			Action	DRS
	1.proxl.perc.intra.single.xml	Xml
	2.proxl.perc.inter.intra.xml	Xml
	20231227_Buchner_Crosslink_01.kojak.txt	Txt
	20231227_Buchner_Crosslink_01.mzXML	Mzxml
	20231227_Buchner_Crosslink_01.perc.intra.pin	Other

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Phosphorylation activates the yeast small heat shock protein Hsp26 by weakening domain contacts in the oligomer ensemble

Project description:Hsp26 is a small heat shock protein (sHsp) from S. cerevisiae. It is known to be activated by dissociation of the oligomer at heat shock temperatures. We wondered whether phosphorylation regulates its activity at physiological temperatures. In Hsp26, 9 phosphorylation sites which are located in different structural elements are known. Our analysis of phospho-mimetic mutations showed that phosphorylation activates Hsp26 at permissive temperature. The cryo-EM structure of the Hsp26 40mer revealed contacts between the conserved core domain of Hsp26 and the so-called thermosensor domain in the N-terminal part of the protein, which are targeted by phosphorylation. Furthermore, the C-terminal extension, which also links subunits within the oligomer, contains several phosphorylation sites. The introduction of negative charges at these strategic positions makes regions in the N-terminal domain accessible for the binding of substrate proteins. Relieving the intrinsic inhibition via different phosphorylation sites allows the fine-tuning of chaperone activity in response to proteotoxic stresses independent of heat activation. The described weakening of domain interactions within and between subunits could be a general regulation principle of sHsps.

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A direct interplay between the co-chaperones Hop and p23 regulates cycle progression and client folding in the Hsp70/Hsp90 chaperone machineries

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The effect of bound RSC/nucleosome complex on chromatin architecture and gene expression at the GAL genes of yeast

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