LIQUID-CHROMATOGRAPHY PURIFICATION AND STUDY OF URIDILYLPOLYNUCLEOTIDE-(5'P-]O)-TYROSINE PHOSPHODIESTERASE

Citation
Mb. Viryasov et al., LIQUID-CHROMATOGRAPHY PURIFICATION AND STUDY OF URIDILYLPOLYNUCLEOTIDE-(5'P-]O)-TYROSINE PHOSPHODIESTERASE, Chromatographia, 45, 1997, pp. 133-137
Citations number
12
Categorie Soggetti
Chemistry Analytical","Biochemical Research Methods
Journal title
ISSN journal
00095893
Volume
45
Year of publication
1997
Supplement
S
Pages
133 - 137
Database
ISI
SICI code
0009-5893(1997)45:<133:LPASOU>2.0.ZU;2-Y
Abstract
LC has been used as a tool for studying uridilylpolynucleotide-(5'P--> O)-phosphodiesteras - an enzyme which hydrolyses specifically the phos phodiester band between picornaviral RNA and viral protein VPg. Accord ing to various chromatographic data, the enzyme forms two types of com plex with nucleic acids: weak ones which dissociate in 200 mM KCl, and others which are stable at concentrations up to 900 mM KCI. 2.5-3-fol d (preparative) or 6-fold (normal scale) purification of the enzyme wa s obtained by size-exclusion chromatography (SEC). Cation-exchange sep aration (4-fold purification) was found to be more suitable as the sec ond enzyme purification step than the earlier anion-exchange method us ed. Three forms of enzyme activity were discovered by hydrophobic-inte raction chromatography on the enzyme preparation obtained by SEC.