Determination of protein transporter function using Raman spectroscopy.

ABSTRACT: Transporter proteins are essential across the tree of life as they provide a cell with a means of exchanging vital metabolites with the external milieu. Characterizing the function of transporters is challenging and traditionally uses methods involving radiolabelled substrates, which requires prolonged exposure times and specialist equipment. Here, we provide an alternative method to the classical uptake assay using Raman spectroscopy to detect the uptake of alkyne-labelled substrates and determine transporter function. As a proof of principle, we demonstrate the method using a candidate nucleotide transporter (ThNTT4) expressed in Escherichia coli, which is shown to transport alkyne-labelled ATP molecules (N6pATP), which was readily detected using Raman spectroscopy. We show that A

SUBMITTER: Gilchrist D 

PROVIDER: S-EPMC12282265 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature