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Humic substances (HS) are the largest constituent of soil organic matter and are considered as a key component of the terrestrial ecosystem. HS may facilitate the transport of organic and inorganic molecules, as well as the sorption interactions with environmentally relevant proteins such as prions. Prions enter the environment through shedding from live hosts, facilitating a sustained incidence of animal prion diseases such as Chronic Wasting Disease and scrapie in cervid and ovine populations, respectively. Changes in prion structure upon environmental exposure may be significant as they can affect prion infectivity and disease pathology. Despite its relevance, the mechanisms of prion interaction with HS are still not completely understood. The goal of this work is to advance a structural-level picture of the encapsulation of recombinant, non-infectious, prion protein (PrP) into different natural HS. We observed that PrP precipitation upon addition of HS is mainly driven by a mechanism of “salting-out” whereby PrP molecules are rapidly removed from the solution and aggregate in insoluble adducts with humic molecules. Importantly, this process does not alter the protein folding since insoluble PrP retains its α-helical content when in complex with HS. The observed ability of HS to promote PrP insolubilization without altering its secondary structure may have potential relevance in the context of “prion ecology”. These results suggest that soil organic matter interacts with prions possibly without altering the protein structures. This may facilitate prions preservation from biotic and abiotic degradation leading to their accumulation in the environment. [ABSTRACT FROM AUTHOR]

Abstract: Knowledge concerning the location of soil enzymes and their kinetics is necessary for a better understanding of their roles in biogeochemical cycles. A rapid dichloromethane fumigation procedure and a new extraction method giving negligible co-extraction of humic substances were used. Fumigation and consequent cell lysis increased the activity of arylsulfatase, but not that of acid and alkaline phosphomonoesterase, suggesting that only the former enzyme was located both extracellularly and intracellularly. Extracted arylsulfatase displayed a Michaelis–Menten constant (K m) significantly lower than in fumigated soil, which in turn was significantly lower than in unfumigated soil. In contrast, each phosphomonoesterase displayed the same K m in extracts, fumigated and unfumigated soils. These results show that activity of soil phosphomonoesterases is not influenced by any direct interaction with soil and, therefore, are more likely to be extracellular enzymes primarily devoted to biogeochemical cycling than are arylsulfatases. [Copyright &y& Elsevier]