Next Article in Journal
Qualitative and Quantitative Analysis of the Major Constituents in WLJ Herbal Tea Using Multiple Chromatographic Techniques
Next Article in Special Issue
Multicellular Human Gastric Cancer Spheroids Mimic the Glycosylation Phenotype of Gastric Carcinomas
Previous Article in Journal
Effect of High Pressure Treatment on Poly(lactic acid)/Nano–TiO2 Composite Films
Previous Article in Special Issue
O-GlcNAc Code’ Mediated Biological Functions of Downstream Proteins
Article Menu
Issue 10 (October) cover image

Export Article

Open AccessArticle
Molecules 2018, 23(10), 2622; https://doi.org/10.3390/molecules23102622

Monitoring Protein Dynamics in Protein O-Mannosyltransferase Mutants In Vivo by Tandem Fluorescent Protein Timers

1
Centre for Organismal Studies (COS), Heidelberg University, 69120 Heidelberg, Germany
2
Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Heidelberg University, 69120 Heidelberg, Germany
3
Deutsches Krebsforschungszentrum (DKFZ), DKFZ-ZMBH Alliance, 69120 Heidelberg, Germany
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 14 September 2018 / Revised: 5 October 2018 / Accepted: 9 October 2018 / Published: 12 October 2018
(This article belongs to the Special Issue Functional Roles of Protein O-Glycosylation)
Full-Text   |   PDF [2146 KB, uploaded 12 October 2018]   |  

Abstract

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. View Full-Text
Keywords: glycosylation; mannosyltransferase; fluorescent protein timers; secretory pathway; O-mannosyl glycans; protein turnover; Saccharomyces cerevisiae; PMT1; PMT2; PMT4; yeast glycosylation; mannosyltransferase; fluorescent protein timers; secretory pathway; O-mannosyl glycans; protein turnover; Saccharomyces cerevisiae; PMT1; PMT2; PMT4; yeast
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material

SciFeed

Share & Cite This Article

MDPI and ACS Style

Castells-Ballester, J.; Zatorska, E.; Meurer, M.; Neubert, P.; Metschies, A.; Knop, M.; Strahl, S. Monitoring Protein Dynamics in Protein O-Mannosyltransferase Mutants In Vivo by Tandem Fluorescent Protein Timers. Molecules 2018, 23, 2622.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top