Development of a Genetically Encoded Sensor for Arginine

The amino acid l-arginine (Arg) plays important roles in multiple metabolic and physiological processes, and changes in its concentration have been implicated in pathological processes. While it is important to measure Arg levels in biological systems directly and in real-time, existing Arg sensors respond to l-ornithine or l-lysine. Here we report ArgS1, a new Arg sensor. It showed a concentration-dependent increase in the ratio Ex488/405 for Arg with an apparent affinity of ∼64 μM and with a dynamic range (ΔR/R0) of 3. ArgS1 responds to Arg in both the cytoplasm and the subcellular organelles. ArgS1 monitored Arg levels in MDA-MB-231 cells, a breast cancer cell line deficient in a key enzyme for Arg synthesis (arginino-succinate synthetase1, ASS1) and amenable to Arg depletion therapy. We found that Arg levels in MDA-MB-231 cells decreased after depletion of extracellular Arg with a concomitant decline in cell viability. When ASS1 was overexpressed in the cells, Arg levels increased and cell viability was also enhanced. Thus, ArgS1 is an effective tool for real-time monitoring of Arg in human cells over a dynamic range of physiological and pathological relevance.