Structure–Functional Examination of Novel Ribonucleoside Hydrolase C from ILimosilactobacillus reuteri/I LR1

• Published in: International journal of molecular sciences Vol. 25; no. 1
• Main Authors: Shaposhnikov, Leonid A, Chikurova, Natalia Yu, Atroshenko, Denis L, Savin, Svyatoslav S, Kleymenov, Sergei Yu, Chernobrovkina, Alla V, Pometun, Evgenii V, Minyaev, Mikhail E, Matyuta, Ilya O, Hushpulian, Dmitry M, Boyko, Konstantin M, Tishkov, Vladimir I, Pometun, Anastasia A
• Format: Journal Article
• Language: English
• Published: MDPI AG 01-12-2023
• Subjects: Analysis; Crystals; Enzymes; Ethylenediaminetetraacetic acid; Hydrolases; Monosaccharides; Scientific equipment and supplies industry; Structure; Sugars; United States; Massachusetts; Germany; Russia
• ISSN: 1422-0067
• DOI: 10.3390/ijms25010538

Ribonucleoside hydrolase C (RihC, EC 3.2.2.1, 3.2.2.2, 3.2.2.3, 3.2.2.7, 3.2.2.8) belongs to the family of ribonucleoside hydrolases Rih and catalyzes the cleavage of ribonucleosides to nitrogenous bases and ribose. RihC is one of the enzymes that are synthesized by lactobacilli in response to the presence of Klebsiella. To characterize this protein from Limosilactobacillus reuteri LR1, we cloned and expressed it. The activity of the enzyme was studied towards a wide range of substrates, including ribonucleosides, deoxyribonucleosides as well as an arabinoside. It was shown that the enzyme is active only with ribonucleosides and arabinoside, with the best substrate being uridine. The thermal stability of this enzyme was studied, and its crystal structure was obtained, which demonstrated the tetrameric architecture of the enzyme and allowed to shed light on a correlation between its structure and enzymatic activity. Comprehensive comparisons of all known RihC structures, both existing crystal structures and computed model structures from various species, were made, allowing for the identification of structural motifs important for enzyme functioning.