Data to "A Semi-Quantitative Yeast Complementation Platform for Characterizing Urea and Ammonia Transport by Membrane Channels"

This repository contains the raw and processed data underlying Figures 2, 3, and 4 in the Current Protocols article: "A Semi-Quantitative Yeast Complementation Platform for Characterizing Urea and Ammonia Transport by Membrane Channels" (DOI: 10.1002/cpz1.70336).

Contents

• Figure 2: Exemplary urea growth curves and pH dependency plots for HpUreI WT, A57C, N-tag+L134C, and the empty vector control. Data include cell counts measured over time at pH 4.0–7.0, normalized urea permeability values, Boltzmann fits, and extracted pKa and Δurea values.
• Figure 3: Ammonia growth assay data for hAQP8, HpUreI, HpUreI E177Q, and the empty vector control, including growth curves in ammonia media (pH 4.0–7.0) and arginine control media, unnormalized and normalized pH dependency plots, averaged relative ammonia permeability (± SEM), and extracted pKa values and ammonia permeability amplitudes.
• Figure 4: Optimization of nitrogen source concentration for urea and ammonia growth assays. Data include normalized permeabilities tested at 0.5, 1.0, 2.0, and 4.0 mM (pH 4.5), demonstrating that 2 mM is the optimal concentration for both solutes.

Data Format

• Spreadsheets (.xlsx or .csv) with time-course cell counts, pH conditions, normalized permeability values, and extracted parameters (pKa, Δurea, permeability amplitudes).

Purpose

This dataset supports the step-by-step protocol for quantifying urea and ammonia permeability and pH-gating of membrane channels using yeast complementation assays. It provides reproducible benchmarks for researchers adapting the method to study other solutes, protein homologs, or mutants.

Usage Notes

• Data are organized by figure and condition.
• Normalization was performed using arginine control media as a reference.
• For questions or collaborations, contact: andreas.horner@jku.at