Comparative Proteome Analysis in Schizosaccharomyces pombe Identifies Metabolic Targets to Improve Protein Production and Secretion

Hung, C. W., Klein, T., Cassidy, L., Linke, D., Lange, S., Anders, U., Bureik, M., Heinzle, E., Schneider, K. and Tholey, Andreas (2016) Comparative Proteome Analysis in Schizosaccharomyces pombe Identifies Metabolic Targets to Improve Protein Production and Secretion Molecular & Cellular Proteomics, 15 (10). pp. 3090-3106. DOI 10.1074/mcp.M115.051474.

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Abstract

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 up-regulation of several amino acid biosyntheses. These amino acids were partially underrepresented in the cellular protein compared with 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, whereas 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. Data are available via ProteomeXchange with identifiers PXD002693 and PXD003016.

Document Type: Article
Additional Information: Times Cited: 0 Hung, Chien-Wen Klein, Tobias Cassidy, Liam Linke, Dennis Lange, Sabrina Anders, Uwe Bureik, Matthias Heinzle, Elmar Schneider, Konstantin Tholey, Andreas
Research affiliation: Kiel University > Kiel Marine Science
OceanRep > The Future Ocean - Cluster of Excellence
Refereed: Yes
DOI etc.: 10.1074/mcp.M115.051474
ISSN: 1535-9476
Projects: Future Ocean
Date Deposited: 26 Feb 2017 11:14
Last Modified: 01 Feb 2018 12:08
URI: http://eprints.uni-kiel.de/id/eprint/36145

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