PXD002693

PXD002693 is an original dataset announced via ProteomeXchange.

Title	Comparative Proteome Analysis in Schizosaccharomyces pombe identifies metabolic targets to improve protein production and secretion
Description	Protein secretion in yeast is a complex process and its efficiency depends on a variety of parameters. We performed a comparative proteome analysis of a set of Schizosaccharomyces pombe strains producing the alpha-glucosidase maltase in increasing amounts to investigate the overall proteomic response of the cell to the burden of protein production along the various steps of protein production and secretion. Proteome analysis of these strains, utilizing an isobaric labeling /two dimensional LC-MALDI MS approach, revealed complex changes, from chaperones and secretory transport machinery to proteins controlling transcription and translation. We also found an unexpectedly high amount of changes in enzyme levels of the central carbon metabolism and a significant upregulation of several amino acid biosyntheses. These amino acids were partially underrepresented in the cellular protein compared to the composition of the model protein. Additional feeding of these amino acids resulted in a 1.5-fold increase in protein secretion. Membrane fluidity was identified as a second bottleneck for high-level protein secretion and addition of fluconazole to the culture caused a significant decrease in ergosterol levels, while protein secretion could be further increased by a factor of 2.1. In summary, we show that high level protein secretion causes global changes of protein expression levels in the cell and that precursor availability and membrane composition limit protein secretion in this yeast. In this respect, comparative proteome analysis is a powerful tool to identify targets for an efficient increase of protein production and secretion in S. pombe. Additional supernatant analysis was performed using LC-ESI MS.
HostingRepository	PRIDE
AnnounceDate	2016-08-08
AnnouncementXML	Submission_2016-08-08_04:14:22.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Andreas Tholey
SpeciesList	scientific name: Schizosaccharomyces pombe; NCBI TaxID: 4896;
ModificationList	iTRAQ4plex-116 reporter+balance reagent acylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive; TOF/TOF 5800

Revision	Datetime	Status	ChangeLog Entry
0	2015-08-10 06:10:48	ID requested
⏵ 1	2016-08-08 04:14:23	announced

Hung CW, Klein T, Cassidy L, Linke D, Lange S, Anders U, Bureik M, Heinzle E, Schneider K, Tholey A, Comparative Proteome Analysis in Schizosaccharomyces pombe Identifies Metabolic Targets to Improve Protein Production and Secretion. Mol Cell Proteomics, 15(10):3090-3106(2016) [pubmed]

curator keyword: Biological

submitter keyword: iTRAQ, 2D-LC MALDI MS, 5800 Proteome Analyzer, isobaric labeling, high/low pH LC

Tholey, Andreas
contact affiliation	Systematic Proteome Research & Bioanalytics, Christian-Albrechts-Univeritaet Kiel, 24105 Kiel, Germany
contact email	a.tholey@iem.uni-kiel.de
lab head
Andreas Tholey
contact affiliation	Systematic Proteome Research & Bioanalytics, University of Kiel
contact email	a.tholey@iem.uni-kiel.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/2016/08/PXD002693

PRIDE project URI

• PRIDE
  1. PXD002693
     1. Label: PRIDE project
     2. Name: Comparative Proteome Analysis in Schizosaccharomyces pombe identifies metabolic targets to improve protein production and secretion