PXD007825

PXD007825 is an original dataset announced via ProteomeXchange.

Title	Quantitative proteomic analysis in Candida albicans using SILAC-based mass spectrometry
Description	Stable isotope labelling by amino acids in cell culture (SILAC) in conjunction with mass spectrometry analysis is a sensitive and reliable technique for quantifying relative differences in protein abundance and post-translational modifications between cell populations. We have developed and utilised SILAC-MS workflows for quantitative proteomics in the fungal pathogen Candida albicans. Arginine metabolism provides important cues for escaping host defences during pathogenesis, which limits the use of auxotrophs in Candida research. Our strategy eliminates the need for engineering arginine auxotrophs for SILAC experiments and allows the use of ARG4 as selectable marker during strain construction. Cells that were auxotrophic for lysine were successfully labelled with both lysine and arginine stable isotopes. We found that prototrophic C. albicans preferentially uses exogenous arginine and downregulates internal production, which allowed it to achieve high incorporation rates. However, similar to other yeast, C. albicans was able to metabolise heavy arginine to heavy proline, which compromised the accuracy of protein quantification. A computational method was developed to correct for the incorporation of heavy proline. In addition, we utilised the developed SILAC labelling in Candida albicans for the global quantitative proteomic analysis of a strain expressing a phosphatase-dead mutant Cdc14PD.
HostingRepository	PRIDE
AnnounceDate	2018-04-17
AnnouncementXML	Submission_2019-03-07_05:43:22.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Iliyana Kaneva
SpeciesList	scientific name: Candida albicans (Yeast); NCBI TaxID: 5476;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2017-09-25 01:31:06	ID requested
1	2018-04-17 00:02:24	announced
⏵ 2	2019-03-07 05:43:24	announced	Updated publication reference for PubMed record(s): 29280593.

Kaneva IN, Longworth J, Sudbery PE, Dickman MJ, Quantitative Proteomic Analysis in Candida albicans Using SILAC-Based Mass Spectrometry. Proteomics, 18(5-6):e1700278(2018) [pubmed]

curator keyword: Biological

submitter keyword: mass spectrometry, Candida albicans, SILAC, Cdc14, nativeSILAC

Mark Dickman
contact affiliation	Department of Chemical and Biological Engineering, University of Sheffield
contact email	m.dickman@sheffield.ac.uk
lab head
Iliyana Kaneva
contact affiliation	University of Sheffield
contact email	iliyana.kaneva@gmail.com
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/2018/04/PXD007825

PRIDE project URI

• PRIDE
  1. PXD007825
     1. Label: PRIDE project
     2. Name: Quantitative proteomic analysis in Candida albicans using SILAC-based mass spectrometry