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While traces are required for microbial utilization, excess amounts are toxic; thus, selenium can be regarded as a biological double-edged sword. Selenium is chemically similar to the essential element sulfur, but curiously, evolution has selected the former over the latter for a subset of oxidoreductases. Enzymes involved in sulfur metabolism are less discriminate in terms of preventing selenium incorporation; however, its specific incorporation into selenoproteins reveals a highly discriminate process that is not completely understood. We have identified SclA, a NifS-like protein in the nosocomial pathogen, Enterococcus faecalis, and characterized its enzymatic activity and specificity for L-selenocysteine over L-cysteine. It is known that Asp-146 is required for selenocysteine specificity in the human selenocysteine lyase. Thus, using computational biology, we compared the bacterial and mammalian enzymes and identified His-100, an Asp-146 ortholog in SclA, and generated site-directed mutants in order to study the residue's potential role in the L-selenocysteine discrimination mechanism. The proteins were overexpressed, purified, and characterized for their biochemical properties. All mutants exhibited varying Michaelis-Menten behavior towards L-selenocysteine, but His-100 was not found to be essential for this activity. Additionally, L-cysteine acted as a competitive inhibitor of all enzymes with higher affinity than L-selenocysteine. Finally, we discovered that SclA exhibited low activity with L-cysteine as a poor substrate regardless of mutations. We conclude that His-100 is not required for L-selenocysteine specificity, underscoring the inherent differences in discriminatory mechanisms between bacterial NifS-like proteins and mammalian selenocysteine lyases. (C) 2021 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.", "subjects" : [ { "otype" : "Classification", "mtid" : 10545, "link" : "/api/classification/10545", "label" : "Biológiai tudományok", "published" : true, "snippet" : true } ], "keywords" : [ { "otype" : "Keyword", "mtid" : 6608, "link" : "/api/keyword/6608", "label" : "enzymology", "published" : true, "oldId" : 6608, "snippet" : true }, { "otype" : "Keyword", "mtid" : 7818, "link" : "/api/keyword/7818", "label" : "Substrate Specificity", "published" : true, "oldId" : 7818, "snippet" : true }, { "otype" : "Keyword", "mtid" : 1008441, "link" : "/api/keyword/1008441", "label" : "Enterococcus faecalis", "published" : true, "oldId" : 1008441, "snippet" : true }, { "otype" : "Keyword", "mtid" : 1576037, "link" : "/api/keyword/1576037", "label" : "Selenocysteine lyase", "published" : true, "snippet" : true }, { "otype" : "Keyword", "mtid" : 1761965, "link" : "/api/keyword/1761965", "label" : "cysteine desulfurase", "published" : true, "snippet" : true }, { "otype" : "Keyword", "mtid" : 2274393, "link" : "/api/keyword/2274393", "label" : "NIFS-LIKE PROTEIN", "published" : true, "snippet" : true } ], "digital" : null, "printed" : null, "sourceYear" : 2021, "foreignEdition" : true, "foreignLanguage" : true, "fullPublication" : true, "conferencePublication" : false, "nationalOrigin" : null, "missingAuthor" : false, "oaType" : "NONE", "oaCheckDate" : "2023-04-27", "oaFree" : false, "citationCount" : 0, "citationCountUnpublished" : 0, "citationCountWoOther" : 0, "independentCitCountWoOther" : 0, "doiCitationCount" : 0, "wosCitationCount" : 0, "scopusCitationCount" : 0, "independentCitationCount" : 0, "unhandledCitationCount" : 0, "citingPubCount" : 0, "independentCitingPubCount" : 0, "unhandledCitingPubCount" : 0, "citedPubCount" : 3, "citedCount" : 3, "ratings" : [ { "otype" : "SjrRating", "mtid" : 11225443, "link" : "/api/sjrrating/11225443", "label" : "sjr:Q1 (2021) Scopus - Medicine (miscellaneous) BIOCHIMIE 0300-9084 1638-6183", "listPos" : 585, "rankValue" : 0.24, "type" : "journal", "ratingType" : { "otype" : "RatingType", "mtid" : 10002, "link" : "/api/ratingtype/10002", "label" : "sjr", "code" : "sjr", "published" : true, "snippet" : true }, "subject" : { "otype" : "ClassificationExternal", "mtid" : 2701, "link" : "/api/classificationexternal/2701", "label" : "Scopus - Medicine (miscellaneous)", "published" : true, "oldId" : 2701, "snippet" : true }, "ranking" : "Q1", "calculation" : "DIRECT", "published" : true, "snippet" : true } ], "ratingsForSort" : "Q1", "hasCitationDuplums" : false, "directInstitutesForSort" : "", "ownerAuthorCount" : 2, "ownerInstituteCount" : 13, "directInstituteCount" : 0, "authorCount" : 4, "contributorCount" : 0, "hasQualityFactor" : true, "link" : "/api/publication/32298275", "label" : "Johnstone Michael A. et al. 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