PXD008812

PXD008812 is an original dataset announced via ProteomeXchange.

Title	The formation of a camalexin-biosynthetic metabolon
Description	Arabidopsis thaliana efficiently synthesizes the antifungal phytoalexin camalexin without apparent release of bioactive intermediates, such as indole-3-acetaldoxime, suggesting channeling of the biosynthetic pathway involving an enzyme complex. To identify such protein interactions, two independent untargeted co-immunoprecipitation (Co-IP) approaches with the biosynthetic enzymes CYP71B15 and CYP71A13 as baits were performed and the camalexin biosynthetic P450 enzymes were co-purified. These interactions were confirmed by targeted Co-IP and förster resonance energy transfer measurements based on fluorescence lifetime imaging (FLIM-FRET). Furthermore, interaction of CYP71A13 and Arabidopsis P450 reductase 1 (ATR1) and recruitment of cytosolic gamma-glutamyl peptidase 1 (GGP1) to the endoplasmic reticulum (ER) was observed. An increased substrate affinity of CYP79B2 in presence of CYP71A13 was shown, indicating substrate channeling by the complex. During camalexin biosynthesis, indol-3-acetonitrile (IAN) is activated by CYP71A13 and glutathionylated to GS-IAN. To clarify whether this is mediated by a specific glutathione transferase (GST), CYP71A13 was expressed together with each of the 54 Arabidopsis GSTs in S. cerevisiae and GS-IAN-synthesis was observed for GSTU2. Corresponding studies of knock-out and overexpression plants did not show an altered camalexin content. However, an influence of the closely related GSTU4 on camalexin biosynthesis was demonstrated. GSTU4 is co-expressed with tryptophan- and camalexin specific enzymes and physically interacts with CYP71A13. Surprisingly, camalexin concentrations were reduced in knock-out and elevated in GSTU4 overexpressing plants. This shows that GSTU4 is not directly involved in camalexin biosynthesis but rather has a regulatory role or is involved in transport or a competing mechanism.
HostingRepository	PRIDE
AnnounceDate	2019-08-26
AnnouncementXML	Submission_2019-08-26_01:30:13.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Stephanie Heinzlmeir
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive; LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2018-01-30 23:32:02	ID requested
⏵ 1	2019-08-26 01:30:14	announced

Dataset with its publication pending

curator keyword: Biological

submitter keyword: camalexin, cytochrome P450

Bernhard Kuster
contact affiliation	Chair of Proteomics and Bioanalytics, Technische Universität München, Germany
contact email	kuster@tum.de
lab head
Stephanie Heinzlmeir
contact affiliation	Chair of Proteomics and Bioanalytics, Technische Universität München, Germany
contact email	stephanie.heinzlmeir@tum.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/08/PXD008812

PRIDE project URI

• PRIDE
  1. PXD008812
     1. Label: PRIDE project
     2. Name: The formation of a camalexin-biosynthetic metabolon