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Protein Modifications in the RER

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Modification of secretory and transmembrane proteins entering the rough ER begins in the ER lumen. These modifications aid in protein folding and stabilize the acquired tertiary structure. Protein modifications in the rough ER co-occur at different stages of protein folding.
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Updated: Jun 18, 2026

Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications

Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications

Published on: October 30, 2013

Leveraging yeast sequestration to study and engineer posttranslational modification enzymes.

Samantha G Martinusen¹, Carl A Denard¹

¹Department of Chemical Engineering, University of Florida, Gainesville, Florida, USA.

Biotechnology and Bioengineering

|December 11, 2023

Summary

This review explores using Saccharomyces cerevisiae organelle sequestration to study and engineer posttranslational modification enzymes (PTM-enzymes). These high-throughput yeast-based methods simplify understanding and reprogramming PTM-enzyme functions.

Keywords:

PTM-enzyme endoplasmic reticulum sequestration protease sortase yeast surface display

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Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications

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Published on: October 30, 2013

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Enzymatic Modification and Flow Cytometry Assessment of Yeast Surface Displayed Proteins

Enzymatic Modification and Flow Cytometry Assessment of Yeast Surface Displayed Proteins

Published on: May 30, 2025

Area of Science:

Biochemistry and Molecular Biology
Synthetic Biology
Biotechnology

Background:

Posttranslational modification enzymes (PTM-enzymes) are crucial for understanding biological processes and disease.
Developing tools to study and engineer PTM-enzymes is essential for advancements in chemical biology, synthetic biology, and biomedicine.
High-throughput functional screens are necessary to decipher PTM-enzyme mechanisms and develop novel functionalities.

Purpose of the Study:

To review the principles and applications of Saccharomyces cerevisiae organelle sequestration for studying and engineering PTM-enzymes.
To highlight the advantages and limitations of different yeast sequestration systems for PTM-enzyme research.
To introduce and discuss yeast endoplasmic reticulum sequestration as a novel approach.

Main Methods:

Discusses outer membrane sequestration using modified yeast surface display.
Explains cytoplasmic sequestration based on enzyme-mediated transcription activation.
Presents a detailed analysis of yeast endoplasmic reticulum sequestration.

Main Results:

Yeast organelle sequestration provides high-throughput methods for PTM-enzyme analysis.
Different sequestration systems offer distinct advantages for measuring and controlling enzyme catalytic efficiencies.
Yeast endoplasmic reticulum sequestration is presented as a new avenue for PTM-enzyme engineering.

Conclusions:

Yeast-based high-throughput sequestration approaches significantly reduce the complexity of studying PTM-enzyme function.
These methods facilitate the reprogramming of PTM-enzymes for improved and novel applications.
Organelle sequestration in Saccharomyces cerevisiae is a powerful strategy for advancing PTM-enzyme research and engineering.