The Hunt for the Closed Conformation of the Fruit-Ripening Enzyme 1-Aminocyclopropane-1-carboxylic Oxidase: A Combined Electron Paramagnetic Resonance and Molecular Dynamics Study

1-Aminocyclopropane-1-carboxylic oxidase (ACCO) is a non-heme iron(II)-containing enzyme involved in the biosynthesis of the phytohormone ethylene, which regulates fruit ripening and flowering in plants. The active conformation of ACCO, and in particular that of the C-terminal part, remains unclear and open and closed conformations have been proposed. In this work, a combined experimental and computational study to understand the conformation and dynamics of the C-terminal part is reported. Site-directed spin-labeling coupled to electron paramagnetic resonance (SDSL-EPR) spectroscopy was used. Mutagenesis experiments were performed to generate active enzymes bearing two paramagnetic labels (nitroxide radicals) anchored on cysteine residues, one in the main core and one in the C-terminal part. Inter-spin distance distributions were measured by pulsed EPR spectroscopy and compared with the results of molecular dynamics simulations. The results reveal the existence of a flexibility of the C-terminal part. This flexibility generates several conformations of the C-terminal part of ACCO that correspond neither to the existing crystal structures nor to the modelled structures. This highly dynamic region of ACCO raises questions on its exact function during enzymatic activity.     

Radionuclide, Metal and Non-metal Levels in Percolated Water from Soils Fertilized with Phosphogypsum

Phosphogypsum (PG) is a by-product of the phosphate fertilizer industry, which is produced by precipitation during the wet process of phosphate rocks. While commercial uses, in agriculture and in manufacturing gypsum board and Portland cement, consume less than a few percent of this by-product, the vast majority is disposed of on land in gypsum. In Brazil, three main industries are responsible for the production and storage of about 5.5 × 10⁶ tons per year. PG may contain trace metals, non-metals, fluorides and natural radionuclides. Since, in Brazil, PG has been used for many years as soil amendment, it is important to know its availability, mainly in aquatic environments used for human consumption. In this case, more restrictive limits must be adopted. This work aimed to evaluate the mobility of metals in sand and clayey soils and, consequently, the contamination of drainage water through greenhouse-scale leaching and transport of toxic elements and radionuclides from soils fertilized with PG to crops. In general, it was observed that elemental concentrations were below the actual detection limit of the equipment for all conditions of interest, indicating a low mobility of the analyzed elements in the soil.     

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