A GC-MS/Single-Cell Method to Evaluate Membrane Transporter Substrate Specificity and Signaling

Overview

Journal Front Mol Biosci

Specialty Biology

Date 2021 Apr 30

PMID 33928121

Citations 14

Authors

Stephen J Fairweather

Shoko Okada

Gregory Gauthier-Coles

Kiran Javed

Angelika Broer

Stefan Broer

Affiliations

Soon will be listed here.

Abstract

Amino acid transporters play a vital role in metabolism and nutrient signaling pathways. Typically, transport activity is investigated using single substrates and competing amounts of other amino acids. We used GC-MS and LC-MS for metabolic screening of oocytes expressing various human amino acid transporters incubated in complex media to establish their comprehensive substrate profiles. For most transporters, amino acid selectivity matched reported substrate profiles. However, we could not detect substantial accumulation of cationic amino acids by SNAT4 and ATB in contrast to previous reports. In addition, comparative substrate profiles of two related sodium neutral amino acid transporters known as SNAT1 and SNAT2, revealed the latter as a significant leucine accumulator. As a consequence, SNAT2, but not SNAT1, was shown to be an effective activator of the eukaryotic cellular growth regulator mTORC1. We propose, that metabolomic profiling of membrane transporters in oocytes can be used to test their substrate specificity and role in intracellular signaling pathways.

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