PXD009033

PXD009033 is an original dataset announced via ProteomeXchange.

Title	Identifying the interacting proteins of lignin biosynthetic P450 enzymes by affinity purification coupled with LC-MS/MS analysis
Description	This project aims to identify the potential interacting proteins of lignin biosynthetic P450 enzymes by affinity purification and LC-MS/MS analysis. To this end, the genomic pieces of three lignin biosynthetic P450 genes C4H, C3’H and F5H were cloned into pMDCpC4H-HPB vector to generate HPB-fusion constructs driven by C4H promoter. The resulted constructs were then respectively transformed into ref3-3 (for C4H), Col-0 WT (for C3'H), and fah1-2 (for F5H) backgrounds and pMDCpC4H-HPB empty vector (as control) was transformed into Col-0 WT to generate transgenic plants. Arabidopsis T2 transgenic plants were grown for 5 weeks in soil and 15 grams of stem tissues were collected for protein extraction. Membrane protein complex was purified by affinity purification using streptavidin beads and subject to LC-MS/MS analysis. Two biological repeats were performed for each construct. For the first repeat (gel-base proteomics), each P450 construct has its control since each experiment was performed separately. For the second repeat (affinity purification,AP-MS), one control was shared for all the three P450 genes.
HostingRepository	PRIDE
AnnounceDate	2018-05-02
AnnouncementXML	Submission_2018-05-23_00:35:22.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Chang-Jun Liu
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2018-02-26 03:17:02	ID requested
1	2018-05-02 00:24:37	announced
⏵ 2	2018-05-23 00:35:23	announced	Updated publication reference for PubMed record(s): 29725099.

Gou M, Ran X, Martin DW, Liu CJ, The scaffold proteins of lignin biosynthetic cytochrome P450 enzymes. Nat Plants, 4(5):299-310(2018) [pubmed]

curator keyword: Biological

submitter keyword: Lignin, C4H, C3'H, F5H, affinity purification, LC-MS/MS

Chang-Jun Liu
contact affiliation	Biology Department Brookhaven National Laboratory Upton, NY 11793
contact email	cliu@bnl.gov
lab head
Chang-Jun Liu
contact affiliation	Brookhaven National Laboratory
contact email	cliu@bnl.gov
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/2018/05/PXD009033

PRIDE project URI

• PRIDE
  1. PXD009033
     1. Label: PRIDE project
     2. Name: Identifying the interacting proteins of lignin biosynthetic P450 enzymes by affinity purification coupled with LC-MS/MS analysis