PXD003016 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Comparative Proteome Analysis in Schizosaccharomyces pombe identifies metabolic targets to improve protein production and secretion |
Description | This dataset contains additional experimental results for the determination of the effect of Fluconazole and amino acid supplementation to the NW10 strain of S. pombe. 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_04:23:07.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 | monohydroxylated residue; iodoacetamide derivatized residue |
Instrument | Q Exactive |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2015-10-07 06:56:20 | ID requested | |
⏵ 1 | 2016-08-08 04:23:08 | announced | |
Publication List
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] |
Keyword List
curator keyword: Biological |
submitter keyword: iTRAQ, 2D-LC MALDI MS, 5800 Proteome Analyzer, isobaric labeling, high/low pH LC |
Contact List
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 | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD003016
- Label: PRIDE project
- Name: Comparative Proteome Analysis in Schizosaccharomyces pombe identifies metabolic targets to improve protein production and secretion