Monitoring protein dynamics in protein O-mannosyltransferase mutants in vivo by tandem fluorescent protein timers
For proteins entering the secretory pathway, a major factor contributing to maturation and homeostasis is glycosylation. One relevant type of protein glycosylation is O-mannosylation, which is essential and evolutionarily-conserved in fungi, animals, and humans. Our recent proteome-wide study in the...
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| Main Authors: | , , , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
12 October 2018
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| In: |
Molecules
Year: 2018, Volume: 23, Issue: 10 |
| ISSN: | 1420-3049 |
| DOI: | 10.3390/molecules23102622 |
| Online Access: | Verlag, Volltext: https://doi.org/10.3390/molecules23102622 Verlag, Volltext: https://www.mdpi.com/1420-3049/23/10/2622 
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| Author Notes: | Joan Castells-Ballester, Ewa Zatorska, Matthias Meurer, Patrick Neubert, Anke Metschies, Michael Knop, Sabine Strahl |
| Summary: | For proteins entering the secretory pathway, a major factor contributing to maturation and homeostasis is glycosylation. One relevant type of protein glycosylation is O-mannosylation, which is essential and evolutionarily-conserved in fungi, animals, and humans. Our recent proteome-wide study in the eukaryotic model organism Saccharomyces cerevisiae revealed that more than 26% of all proteins entering the secretory pathway receive O-mannosyl glycans. In a first attempt to understand the impact of O-mannosylation on these proteins, we took advantage of a tandem fluorescent timer (tFT) reporter to monitor different aspects of protein dynamics. We analyzed tFT-reporter fusions of 137 unique O-mannosylated proteins, mainly of the secretory pathway and the plasma membrane, in mutants lacking the major protein O-mannosyltransferases Pmt1, Pmt2, or Pmt4. In these three pmtΔ mutants, a total of 39 individual proteins were clearly affected, and Pmt-specific substrate proteins could be identified. We observed that O-mannosylation may cause both enhanced and diminished protein abundance and/or stability when compromised, and verified our findings on the examples of Axl2-tFT and Kre6-tFT fusion proteins. The identified target proteins are a valuable resource towards unraveling the multiple functions of O-mannosylation at the molecular level. |
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| Item Description: | Gesehen am 02.04.2019 |
| Physical Description: | Online Resource |
| ISSN: | 1420-3049 |
| DOI: | 10.3390/molecules23102622 |