RESUMO
Hydrogen sulfide (H2S) is an important gasotransmitter in both animals and plants. Many physiological events, including responses to stress, have been suggested to involve H2S, at least in part. On the other hand, numerous responses have been reported following treatment with H2S, including changes in the levels of antioxidants and the activities of transcription factors. Therefore, it is important to understand and unravel the events that are taking place downstream of H2S in signaling pathways. H2S is known to interact with other reactive signaling molecules such as reactive oxygen species (ROS) and nitric oxide (NO). One of the mechanisms by which ROS and NO have effects in a cell is the modification of thiol groups on proteins, by oxidation or S-nitrosylation, respectively. Recently, it has been reported that H2S can also modify thiols. Here we report a method for the determination of thiol modifications on proteins following the treatment with biological samples with H2S donors. Here, the nematode Caenorhabditis elegans is used as a model system but this method can be used for samples from other animals or plants.
Assuntos
Caenorhabditis elegans/efeitos dos fármacos , Proteínas de Helminto/isolamento & purificação , Sulfeto de Hidrogênio/farmacologia , Processamento de Proteína Pós-Traducional , Compostos de Sulfidrila/metabolismo , Animais , Comportamento Animal/efeitos dos fármacos , Caenorhabditis elegans/metabolismo , Dissulfetos/química , Dissulfetos/metabolismo , Eletroforese em Gel de Poliacrilamida , Fluoresceínas/química , Proteínas de Helminto/química , Proteínas de Helminto/metabolismo , Peróxido de Hidrogênio/farmacologia , Sulfeto de Hidrogênio/metabolismo , Movimento/efeitos dos fármacos , Óxido Nítrico/metabolismo , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Coloração e Rotulagem/métodos , Compostos de Sulfidrila/química , Sulfetos/química , Sulfetos/farmacologiaRESUMO
Effects of hydrogen sulfide (H(2)S) on plant physiology have been previously studied, but such studies have relied on the use of NaSH as a method for supplying H(2)S to tissues. Now new compounds which give a less severe H(2)S shock and a more prolonged exposure to H(2)S have been developed. Here the effects of one such compound, GYY4137, has been investigated to determine its effects on stomatal closure in Arabidopsis thaliana. It was found that both NaSH and GYY4137 caused stomatal opening in the light and prevented stomatal closure in the dark. Nitric oxide (NO) has been well established as a mediator of stomatal movements and here it was found that both NaSH and GYY4137 reduced the accumulation of NO in guard cells, perhaps suggesting a mode of action for H(2)S in this system. GYY4137, and future related compounds, will be important tools to unravel the effects of plant exposure to H(2)S and to determine how H(2)S may fit into plant cell signalling pathways.
