Počet záznamů: 1
Variations in the enzymatic activity of S1-type nucleases results from differences in their active site structures
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SYSNO ASEP 0584258 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Variations in the enzymatic activity of S1-type nucleases results from differences in their active site structures Tvůrce(i) Krela, Rafal (BC-A) RID, ORCID
Poreba, M. (PL)
Lesniewicz, K. (PL)Celkový počet autorů 3 Číslo článku 130424 Zdroj.dok. Biochimica et Biophysica Acta-General Subjects. - : Elsevier - ISSN 0304-4165
Roč. 1867, č. 10 (2023)Poč.str. 9 s. Jazyk dok. eng - angličtina Země vyd. NL - Nizozemsko Klíč. slova Non-zinc-dependent activity ; S1-like nuclease ; Active-site ; Acanthamoeba castellanii ; Physcomitrella patens Vědní obor RIV EB - Genetika a molekulární biologie Obor OECD Biochemistry and molecular biology Způsob publikování Open access Institucionální podpora BC-A - RVO:60077344 UT WOS 001058918800001 EID SCOPUS 85165714520 DOI 10.1016/j.bbagen.2023.130424 Anotace Background: S1-like nucleases are widespread enzymes commonly used in biotechnology and molecular biology. Although it is commonly believed that they are mainly Zn2+ dependent acidic enzymes, we have found that numerous members of this family deviate from this rule. Therefore, in this work, we decided to check how broad is the range of non-zinc-dependent S1-like nucleases and what is the molecular basis of their activities. Methods: S1-like nucleases chosen for analysis were achieved through heterologous expression in appropriate eukaryotic hosts. To characterize nucleases active-site properties, point mutations were introduced in selected positions. The enzymatic activities of wild-type and mutant nucleases were tested by in-gel nuclease activity assay. Results: We discovered that S1-like nucleases encoded by non-vascular plants and single-celled protozoa, like their higher plant homologues, exhibit a large variety of catalytic properties. We have shown that these individual properties are determined by specific non-conserved active site residues. Conclusions: Our findings demonstrate that mutations that occur during evolution can significantly alter the catalytic properties of S1-like nucleases. As a result, different ions can compete for particular S1-type nucleases active sites. This phenomenon undermines the existing classification of S1-like nucleases. General significance: Our findings have numerous implications for applications and understanding the S1-like nucleases biological functions. For example, new biotechnological applications should take into account their unexpected catalytic properties. Moreover, these results demonstrate that the trinuclear zinc-based model commonly used to characterize the catalytic activities of S1-like nucleases is insufficient to explain the actions of non-zinc-dependent members of this family. Pracoviště Biologické centrum (od r. 2006) Kontakt Dana Hypšová, eje@eje.cz, Tel.: 387 775 214 Rok sběru 2024 Elektronická adresa https://www.sciencedirect.com/science/article/pii/S0304416523001228?via%3Dihub
Počet záznamů: 1