Characterization of inosine-uridine nucleoside hydrolase (RihC) from Escherichia coli.

Autor:	Arivett B; Department of Biology, Middle Tennessee State University, Murfreesboro, TN 37132, USA., Farone M; Department of Biology, Middle Tennessee State University, Murfreesboro, TN 37132, USA., Masiragani R; Department of Chemistry, Middle Tennessee State University, Murfreesboro, TN 37132, USA., Burden A; Department of Chemistry, Middle Tennessee State University, Murfreesboro, TN 37132, USA., Judge S; Department of Chemistry, Middle Tennessee State University, Murfreesboro, TN 37132, USA., Osinloye A; Department of Chemistry, Middle Tennessee State University, Murfreesboro, TN 37132, USA., Minici C; Division of Immunology, Transplantation and Infectious Diseases, Scientific Institute San Raffaele, Milan Italy., Degano M; Division of Immunology, Transplantation and Infectious Diseases, Scientific Institute San Raffaele, Milan Italy., Robinson M; Department of Chemistry, Middle Tennessee State University, Murfreesboro, TN 37132, USA., Kline P; Department of Chemistry, Middle Tennessee State University, Murfreesboro, TN 37132, USA. Electronic address: paul.kline@mtsu.edu.
Jazyk:	angličtina
Zdroj:	Biochimica et biophysica acta [Biochim Biophys Acta] 2014 Mar; Vol. 1844 (3), pp. 656-62. Date of Electronic Publication: 2014 Jan 25.
DOI:	10.1016/j.bbapap.2014.01.010
Abstrakt:	A non-specific nucleoside hydrolase from Escherichia coli (RihC) has been cloned, overexpressed, and purified to greater than 95% homogeneity. Size exclusion chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis show that the protein exists as a homodimer. The enzyme showed significant activity against the standard ribonucleosides with uridine, xanthosine, and inosine having the greatest activity. The Michaelis constants were relatively constant for uridine, cytidine, inosine, adenosine, xanthosine, and ribothymidine at approximately 480μM. No activity was exhibited against 2'-OH and 3'-OH deoxynucleosides. Nucleosides in which additional groups have been added to the exocyclic N6 amino group also exhibited no activity. Nucleosides lacking the 5'-OH group or with the 2'-OH group in the arabino configuration exhibited greatly reduced activity. Purine nucleosides and pyrimidine nucleosides in which the N7 or N3 nitrogens respectively were replaced with carbon also had no activity. (Copyright © 2014 Elsevier B.V. All rights reserved.)
Databáze:	MEDLINE
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