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Uptake Assay for Radiolabeled Peptides in Yeast   

Edited by
Yanjie Li
Reviewed by
Anonymous reviewer
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Original research article

A brief version of this protocol appeared in:
Eukaryotic Cell
Oct 2007

Abstract

We describe an assay for measuring the uptake of radioactive peptides into the yeast Saccharomyces cerevisiae. The methods presented here can be adapted to measure a variety of substrates transported into any bacterial or fungal cell via specific carrier-mediated systems.

Background

Di/tripeptides and larger oligopeptides are sources of amino acids for protein synthesis, or may serve as carbon and nitrogen precursors for energy production and biosynthesis of metabolites in all organisms. Uptake of di/tripeptides and oligopeptides across the cell membrane is facilitated by peptide transporters. Measuring the accumulation of radiolabeled peptide provides an experimental approach to determine uptake across a specific peptide transport protein in a variety of cell types, including the yeast Saccharomyces cerevisiae. Appropriate experimental design allows for the determination of kinetic parameters such as the affinity and capacity of system under investigation. Dipeptides, such as radiolabeled Leu-Leu, can be used to measure transport across the di/tripeptide transporter Ptr2p of S. cerevisiae. We use the transport of dipeptides across Ptr2p in yeast as an example.

Copyright: © 2016 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Hauser, M., Cai, H., Naider, F. and Becker, J. M. (2016). Uptake Assay for Radiolabeled Peptides in Yeast. Bio-protocol 6(22): e2026. DOI: 10.21769/BioProtoc.2026.
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