Journal
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS
Volume 1841, Issue 2, Pages 215-226Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbalip.2013.11.005
Keywords
Pichia pastoris; Endoplasmic reticulum; Proteome; Sphingolipids; Phospholipid; Fatty acid
Funding
- Austrian Science Fund FWF [TRP 9]
- Federal Ministry of Economy, Family and Youth (BMWFJ)
- Federal Ministry of Traffic, Innovation and Technology (bmvit)
- Styrian Business Promotion Agency SFG
- Standortagentur Tirol
- ZIT - Technology Agency of the City of Vienna through the COMET
- Microbiology and Biochemistry program of the Goettingen Graduate School of Neurosciences and Molecular Biology (GGNB)
- Austrian Science Fund (FWF) [TRP 9] Funding Source: researchfish
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The methylotrophic yeast Pichia pastoris is a popular yeast expression system for the production of heterologous proteins in biotechnology. Interestingly, cell organelles which play an important role in this process have so far been insufficiently investigated. For this reason, we started a systematic approach to isolate and characterize organelles from P. pastoris. In this study, we present a procedure to isolate microsomal membranes at high purity. These samples represent endoplasmic reticulum (ER) fractions which were subjected to molecular analysis of lipids and proteins. Organelle lipidomics included a detailed analysis of glycerophospholipids, fatty acids, sterols and sphingolipids. The microsomal proteome analyzed by mass spectrometry identified typical proteins of the ER known from other cell types, especially Saccharomyces cerevisiae, but also a number of unassigned gene products. The lipidome and proteome analysis of P. pastoris microsomes are prerequisite for a better understanding of functions of this organelle and for modifying this compartment for biotechnological applications. (C) 2013 Elsevier B.V. All rights reserved.
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