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Article: Angiostrongylus cantonensis: Characterization of Thymidylate Synthetase

TitleAngiostrongylus cantonensis: Characterization of Thymidylate Synthetase
Authors
Issue Date1994
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/yexpr
Citation
Experimental Parasitology, 1994, v. 79 n. 4, p. 526-535 How to Cite?
AbstractThymidylate synthetase (TS) is the only enzyme that catalyzes the formation of thymidine nucleotides in Angiostrongylus cantonensis. A fraction enriched in TS was obtained from the gravid nematode by gel filtration and affinity chromatography using methotrexate-agarose. TS, which was well separated from dihydrofolate reductase, has a relative molecular mass of 66 kDa. By electrophoresis in sodium dodecyl sulphate gel, a major protein band corresponding to 31 kDa was observed. This band was shown to be TS by comparing the electrophoretic mobility with an enzyme preparation bound with [6-3H]5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP). Therefore, the enzyme is composed of two identical or very similar subunits. Velocity studies and product inhibition patterns revealed that the TS reaction undergoes a sequential mechanism in which 2'-deoxyuridine 5'-monophosphate (dUMP) is the first substrate added to the active site and thymidine 5'-monophosphate is the last product released. The apparent Km values for dUMP and 5,10-methylenetetrahydrofolate are 10 and 185 microM, respectively. FdUMP and trimethoprim inhibited the parasite TS competitively with dUMP and the Ki values of 23.5 nM and 852 microM, respectively. Methotrexate was a noncompetitive inhibitor of TS. At 0.2 mM 5,10-methylenetetrafolate, 1 mM methotrexate inhibited the activity by 74%.
Persistent Identifierhttp://hdl.handle.net/10722/223064
ISSN
2015 Impact Factor: 1.623
2015 SCImago Journal Rankings: 0.791

 

DC FieldValueLanguage
dc.contributor.authorSo, NN-
dc.contributor.authorWong, PCL-
dc.contributor.authorKo, RCC-
dc.date.accessioned2016-02-18T00:52:27Z-
dc.date.available2016-02-18T00:52:27Z-
dc.date.issued1994-
dc.identifier.citationExperimental Parasitology, 1994, v. 79 n. 4, p. 526-535-
dc.identifier.issn0014-4894-
dc.identifier.urihttp://hdl.handle.net/10722/223064-
dc.description.abstractThymidylate synthetase (TS) is the only enzyme that catalyzes the formation of thymidine nucleotides in Angiostrongylus cantonensis. A fraction enriched in TS was obtained from the gravid nematode by gel filtration and affinity chromatography using methotrexate-agarose. TS, which was well separated from dihydrofolate reductase, has a relative molecular mass of 66 kDa. By electrophoresis in sodium dodecyl sulphate gel, a major protein band corresponding to 31 kDa was observed. This band was shown to be TS by comparing the electrophoretic mobility with an enzyme preparation bound with [6-3H]5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP). Therefore, the enzyme is composed of two identical or very similar subunits. Velocity studies and product inhibition patterns revealed that the TS reaction undergoes a sequential mechanism in which 2'-deoxyuridine 5'-monophosphate (dUMP) is the first substrate added to the active site and thymidine 5'-monophosphate is the last product released. The apparent Km values for dUMP and 5,10-methylenetetrahydrofolate are 10 and 185 microM, respectively. FdUMP and trimethoprim inhibited the parasite TS competitively with dUMP and the Ki values of 23.5 nM and 852 microM, respectively. Methotrexate was a noncompetitive inhibitor of TS. At 0.2 mM 5,10-methylenetetrafolate, 1 mM methotrexate inhibited the activity by 74%.-
dc.languageeng-
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/yexpr-
dc.relation.ispartofExperimental Parasitology-
dc.subject.meshAngiostrongylus cantonensis - enzymology-
dc.subject.meshThymidylate synthase - antagonists & inhibitors - isolation & purification - metabolism-
dc.subject.meshMethotrexate - pharmacology-
dc.subject.meshChemical fractionation-
dc.subject.meshChromatography, affinity-
dc.titleAngiostrongylus cantonensis: Characterization of Thymidylate Synthetase-
dc.typeArticle-
dc.identifier.emailKo, RCC: rcko@hkucc.hku.hk-
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1006/expr.1994.1113-
dc.identifier.pmid8001663-
dc.identifier.scopuseid_2-s2.0-0028556515-
dc.identifier.hkuros1941-
dc.identifier.volume79-
dc.identifier.issue4-
dc.identifier.spage526-
dc.identifier.epage535-

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