Protocol to evaluate translation inhibition in Candida species using fluorescence microscopy and flow cytometry.

Translation inhibitors have therapeutic potential against Candida species. Here, we present a protocol to measure translation inhibition in Candida spp. We describe steps for employing an alkynylated methionine analog, L-homopropargylglycine (HPG), that becomes incorporated into newly synthesized proteins. We then detail procedures to perform a click reaction of the alkyne with a fluorescent azide, which is visualized using fluorescence microscopy and quantified by flow cytometry. For complete details on the use and execution of this protocol, please refer to Puumala et al., ¹ Fu et al., ² and Iyer et al. ³ .
Competing Interests: Declaration of interests L.E.C. is a co-founder and shareholder in Bright Angel Therapeutics, a platform company for development of novel antifungal therapeutics. L.E.C. is a Science Advisor for Kapoose Creek, a company that harnesses the therapeutic potential of fungi.
(Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)