Glycolytic Proteins Interact With Intracellular Melatonin in Saccharomyces cerevisiae

Abstract

Melatonin is a bioactive compound that is present in fermented beverages and synthesized by yeast during alcoholic fermentation. Many studies have shown that melatonin interacts with some mammalian proteins, such as sirtuins or orphan receptor family proteins. The aim of this study was to determine the intracellular synthesis profile of melatonin in Saccharomyces cerevisiae and to identify the proteins that may interact with this molecule in yeast cells. Melatonin from fermentation samples was analyzed by liquid chromatography mass spectrometry, and proteins bound to melatonin were immunopurified by melatonin-IgG-Dynabeads. Melatonin was produced intracellularly in the lag phase of yeast growth and was exported to the extracellular media during the stationary phase. During this period, melatonin was bound to six proteins with molecular weights from 55 to 35 kDa. Sequence analysis showed that most proteins shared high levels of homology with glycolytic enzymes. An RNA-binding protein was also identified, the elongation alpha factor, which is related to mitochondria. This study reports for the first time the interaction of melatonin and proteins inside yeast cells. These results highlight the possible role of melatonin as a signal molecule and provide a new perspective for understanding its role in yeast.

Melatonin in yeast and fermented beverages: analytical tools for detection, physiological role and biosynthesis

“…A screening of Saccharomyces and non-Saccharomyces strains revealed that melatonin synthesis took place at the end of the exponential growth phase, and its presence during fermentation followed a zigzag pattern that appeared and disappeared (59,78). These results were also corroborated during grape must fermentation (79).…”

Transcriptomic Insights into the Effect of Melatonin in Saccharomyces cerevisiae in the Presence and Absence of Oxidative Stress

“…Indeed, it has been reported that hypoxia induces the formation of foci containing glycolytic enzymes, which increases the incorporation of carbon into pyruvate and oxaloacetate [ 91 ]. Recently, melatonin has been shown to bind a complex of glycolytic enzymes [ 17 ], and this binding would be related to yeast fermentative capacity [ 18 ]. Several glycolytic genes found in this complex, such as PYC2 and TDH3 , were upregulated by melatonin in non-stressed cells ( Figure 1 B,E, Dataset S1 ).…”

“…For intracellular melatonin quantification, 10 8 cells were harvested from the samples at 0 h (just before stress induction) and 1 h after the stress induction by centrifugation at 4700× g for 15 min. Intracellular melatonin quantification was performed as previously described by Morcillo-Parra et al [ 17 ]. In brief, melatonin was extracted by adapting the boiling buffered ethanol method described by Gonzalez et al [ 28 ] and quantified by liquid chromatography mass spectrometry using a liquid chromatograph coupled to a triple quadrupole mass spectrometer (LC-MS/MS Agilent G6410A; Agilent Technologies Inc., Santa Clara, CA, USA).…”

“…Recent studies have demonstrated a protective function of melatonin against oxidative stress and UV radiation [ 14 , 15 , 16 ] and indicated a hypothetical role as a signaling molecule [ 7 ]. The recent finding that melatonin interacts with glycolytic proteins during alcoholic fermentation reinforces the latter role, which seems to be linked to fermentative metabolism [ 17 , 18 ].…”

Naturally occurring melatonin: Sources and possible ways of its biosynthesis

“…More recently, it was found that glycolytic proteins may have interaction with intracellular MEL in S . cerevisiae and thereby restrict the recovery of this molecule during the first 24 h of lag phase from fermented products (Morcillo‐Parra, Valera, Beltran, Mas, & Torija, 2019). However, after this time the MEL that has been produced intracellularly in the lag phase begins to be exported to extracellular media during the stationary phase.…”