PXD013494

PXD013494 is an original dataset announced via ProteomeXchange.

Title	In vivo trapping of proteins interacting with the chloroplast CLPC1 chaperone
Description	The chloroplast stromal CLP protease system is essential for growth and development. It consists of a proteolytic CLP core complex that likely dynamically interacts with oligomeric rings of CLPC1, CLPC2 or CLPD AAA+ chaperones. These ATP-dependent chaperones are predicted to bind and unfold CLP protease substrates, frequently aided by adaptors (recognins), and feed them into the proteolytic CLP core for degradation. To identify new substrates and possible also new adaptors for the chloroplast CLP protease system, we generated an in vivo CLPC1 substrate trap with a C-terminal STREPII affinity tag in Arabidopsis thaliana by mutating critical glutamate residues (E374A and E718A) in the two Walker B domains of CLPC1 required for hydrolysis of ATP (CLPC1-TRAP). Based on homology to non-plant CLPB/C chaperones, it is predicted that interacting substrates are unable to be released, i.e. they are trapped. When expressed in wild-type, this CLPC1-TRAP induced a dominant visible phenotype, whereas no viable mutants that express CLPC1-TRAP in the clpc1-1 null mutant could be recovered. Affinity purification of the CLPC1-TRAP resulted in a dozen proteins highly enriched compared to affinity purified CLPC1 with a C-terminal STREPII affinity tag (CLPC1-WT). These enriched proteins likely represent CLP protease substrates and/or new adaptors. Several of these trapped proteins over-accumulated in clp mutants and/or were found as interactions for the adaptor CLPS1, supporting their functional relationship to CLP function. Importantly, affinity purification of this CLPC1-TRAP also showed high enrichment of all CLPP, CLPR and CLPT subunits, indicating stabilization of the CLPC to CLP core interaction and providing direct support for their physical and functional interaction.
HostingRepository	PRIDE
AnnounceDate	2019-05-14
AnnouncementXML	Submission_2019-05-14_06:44:45.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD013494
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Giulia Friso
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	monohydroxylated residue; deaminated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2019-04-15 01:27:45	ID requested
⏵ 1	2019-05-14 06:44:47	announced

Montandon C, Friso G, Liao JR, Choi J, van Wijk KJ, In Vivo Trapping of Proteins Interacting with the Chloroplast CLPC1 Chaperone: Potential Substrates and Adaptors. J Proteome Res, 18(6):2585-2600(2019) [pubmed]

submitter keyword: In vivo trapping-proteins interaction- chloroplast - CLPC1 chaperone

Klaas van Wijk
contact affiliation	Cornell University, Plant Biology section
contact email	kv35@cornell.edu
lab head
Giulia Friso
contact affiliation	Plant Biology
contact email	gf32@cornell.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD013494
     1. Label: PRIDE project
     2. Name: In vivo trapping of proteins interacting with the chloroplast CLPC1 chaperone