Cargando…

The bud tip is the cellular hot spot of protein secretion in yeasts

Yeasts are valuable hosts for recombinant protein production. Among them, Pichia pastoris is frequently used for production of secreted proteins, and much effort was made to improve the secretion efficiency of this expression platform. However, the knowledge on the secretion machinery is mainly base...

Descripción completa

Detalles Bibliográficos
Autores principales: Puxbaum, Verena, Gasser, Brigitte, Mattanovich, Diethard
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4989006/
https://www.ncbi.nlm.nih.gov/pubmed/27338576
http://dx.doi.org/10.1007/s00253-016-7674-6
_version_ 1782448497188929536
author Puxbaum, Verena
Gasser, Brigitte
Mattanovich, Diethard
author_facet Puxbaum, Verena
Gasser, Brigitte
Mattanovich, Diethard
author_sort Puxbaum, Verena
collection PubMed
description Yeasts are valuable hosts for recombinant protein production. Among them, Pichia pastoris is frequently used for production of secreted proteins, and much effort was made to improve the secretion efficiency of this expression platform. However, the knowledge on the secretion machinery is mainly based on studies in Saccharomyces cerevisiae. Therefore, it is of great interest for targeted improvement of the system to learn more about the secretion process in P. pastoris. Using human serum albumin, a protein which is produced in high quantities in P. pastoris, we show here the secretion pathway of this protein. During passage of the secretory route, the recombinant protein is mainly localized in the endoplasmic reticulum (ER) and in COPII vesicles, and is inherited to the daughter cell via the perinuclear ER. The final release to the cell exterior occurs at the bud, initiating at the bud tip and later spreading over the entire bud surface. The same polarized secretion pattern was observed for a recombinant antibody light chain and the native secretory protein Epx1 of P. pastoris. Clarifying the point of release of secretory proteins will have major impact on engineering the secretory pathway of P. pastoris and other budding yeasts. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00253-016-7674-6) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4989006
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Springer Berlin Heidelberg
record_format MEDLINE/PubMed
spelling pubmed-49890062016-09-01 The bud tip is the cellular hot spot of protein secretion in yeasts Puxbaum, Verena Gasser, Brigitte Mattanovich, Diethard Appl Microbiol Biotechnol Applied Microbial and Cell Physiology Yeasts are valuable hosts for recombinant protein production. Among them, Pichia pastoris is frequently used for production of secreted proteins, and much effort was made to improve the secretion efficiency of this expression platform. However, the knowledge on the secretion machinery is mainly based on studies in Saccharomyces cerevisiae. Therefore, it is of great interest for targeted improvement of the system to learn more about the secretion process in P. pastoris. Using human serum albumin, a protein which is produced in high quantities in P. pastoris, we show here the secretion pathway of this protein. During passage of the secretory route, the recombinant protein is mainly localized in the endoplasmic reticulum (ER) and in COPII vesicles, and is inherited to the daughter cell via the perinuclear ER. The final release to the cell exterior occurs at the bud, initiating at the bud tip and later spreading over the entire bud surface. The same polarized secretion pattern was observed for a recombinant antibody light chain and the native secretory protein Epx1 of P. pastoris. Clarifying the point of release of secretory proteins will have major impact on engineering the secretory pathway of P. pastoris and other budding yeasts. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00253-016-7674-6) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2016-06-23 2016 /pmc/articles/PMC4989006/ /pubmed/27338576 http://dx.doi.org/10.1007/s00253-016-7674-6 Text en © The Author(s) 2016 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Applied Microbial and Cell Physiology
Puxbaum, Verena
Gasser, Brigitte
Mattanovich, Diethard
The bud tip is the cellular hot spot of protein secretion in yeasts
title The bud tip is the cellular hot spot of protein secretion in yeasts
title_full The bud tip is the cellular hot spot of protein secretion in yeasts
title_fullStr The bud tip is the cellular hot spot of protein secretion in yeasts
title_full_unstemmed The bud tip is the cellular hot spot of protein secretion in yeasts
title_short The bud tip is the cellular hot spot of protein secretion in yeasts
title_sort bud tip is the cellular hot spot of protein secretion in yeasts
topic Applied Microbial and Cell Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4989006/
https://www.ncbi.nlm.nih.gov/pubmed/27338576
http://dx.doi.org/10.1007/s00253-016-7674-6
work_keys_str_mv AT puxbaumverena thebudtipisthecellularhotspotofproteinsecretioninyeasts
AT gasserbrigitte thebudtipisthecellularhotspotofproteinsecretioninyeasts
AT mattanovichdiethard thebudtipisthecellularhotspotofproteinsecretioninyeasts
AT puxbaumverena budtipisthecellularhotspotofproteinsecretioninyeasts
AT gasserbrigitte budtipisthecellularhotspotofproteinsecretioninyeasts
AT mattanovichdiethard budtipisthecellularhotspotofproteinsecretioninyeasts