2′5′ Obgoadenylate synthetase, an interferon induced enzyme: direct assay methods for the products, 2′5′ ohgoadenylates and 2′5′ co-oligonucleotides

Justesen, Just; Ferbus, Didier; Thang, M.N.

doi:10.1093/nar/8.14.3073

Cited by 64 publications

(24 citation statements)

References 12 publications

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“…33 P]ATP with subsequent separation of the radioactive 2-5A products by thin layer chromatography on polyethyleneimine plates (21) (Fig. 6).…”

Section: Resultsmentioning

confidence: 99%

“…Insect cells in 96-well microtiter plates were infected with serial dilutions of transfection solution, and cellular DNA was then examined for the presence of the 2-5A synthetase gene by dot-blot analysis (20). Recombinant baculoviruses were tested for their ability to produce 2-5A synthetase by an enzymatic activity test (21).…”

Section: Baculovirus Expression Of Human 2ј-5јmentioning

confidence: 99%

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Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Hartmann¹,

Nørby²,

Martensen³

et al. 1998

Journal of Biological Chemistry

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A number of small RNA molecules that are high affinity ligands for the 46-kDa form of human 2-5 oligoadenylate synthetase have been identified by the SELEX method. Surface plasmon resonance analysis indicates that these RNAs bind to the enzyme with dissociation constants in the nanomolar range. Competition experiments indicate that the binding site for the small RNAs on the 2-5 oligoadenylate synthetase molecule at least partially overlaps that for the synthetic doublestranded RNA, poly(I)⅐poly(C). Several of the RNAs function as potent activators of 2-5 oligoadenylate synthetase in vitro, although there is no correlation between binding affinity and ability to activate. The RNA aptamers having the strongest activation potential appear to have few base-paired regions. This suggests that 2-5 oligoadenylate synthetase, which has previously been believed to be activated only by double-stranded RNA, can also be activated by RNA ligands with little secondary structure. Since 2-5 oligoadenylate synthetase possesses no homology to other known RNA-binding proteins, the development of small specific ligands by SELEX should facilitate studies of RNA-protein interactions and may reveal novel features of the structurefunction relationships involving this enzyme. Double-stranded RNA (dsRNA) 1 binds to and activates a number of interferon-induced proteins, namely the family of 2Ј-5Ј oligoadenylate synthetases (2-5A synthetase) and the protein kinase PKR (reviewed in Refs.

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“…33 P]ATP with subsequent separation of the radioactive 2-5A products by thin layer chromatography on polyethyleneimine plates (21) (Fig. 6).…”

Section: Resultsmentioning

confidence: 99%

Section: Baculovirus Expression Of Human 2ј-5јmentioning

confidence: 99%

Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Hartmann¹,

Nørby²,

Martensen³

et al. 1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Cell lysates were pelleted for 6 min in an Eppendorf centrifuge and the cell extracts were stored at 7808C until assayed. For the 2'5'A synthetase assay, 100 ml of each cell extract were tested as described (Justesen et al, 1980). One unit of 2'5'A synthetase activity corresponds to 1 nmol ATP converted/h at 378C and its speci®c activity is expressed as units/mg of protein in the cell extract.…”

Section: Assay Of 2'5'a Synthetasementioning

confidence: 99%

Retinoic acid enhances the expression of interferon-induced proteins: evidence for multiple mechanisms of action

et al. 1997

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“…1) biological assays (RNA-cleavage assay (21-23), protein synthesis Inhibition assay (24)) 2) binding assays (radiobinding assay (l 5), radioimmunoassay (25), enzymeimmunoassay (26,27)) and 3) assays that determine 2 / -5'-oligoadenylates formed during the 2 / -5'-oligoadenylate synthetase reaction (14,16,17,28,29).…”

Section: Discussiopmentioning

confidence: 99%

Methods for the Determination of the Interferon-Induced Enzyme 2'-5' Oligoadenylate Synthetase in Mononuclear Blood Cells

Bruchelt¹,

Beck²,

Schilbach-Stückle³

et al. 1987

Clinical Chemistry and Laboratory Medicine

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Summary:The determination of 2'-5'-oligoadenylate synthetase in peripheral blood mononuclear cells is used äs a biological response parameter during therapy with interferon and in the diagnosis of diseases related to the interferon System. In this communication, some general aspects concerning the preparation of 2'-5'-oligoadenylate synthetase from peripheral blood mononuclear cells and the incubation conditions of the 2'-5'-oligoadenylate synthetase reaction are reported. Four analytical procedures for the determination of the products formed during the 2'-5'-oligoadenylate synthetase reaction were comparatively investigated and the advantages and limitations of the assays are discussed. As an example of possible clinical application, the levels of 2'-5'-oligoadenylate synthetase were determined in the mononuclear cell fraction of peripheral blood from healthy persons äs well äs from children with chronic myelogenous leukaemia.

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2′5′ Obgoadenylate synthetase, an interferon induced enzyme: direct assay methods for the products, 2′5′ ohgoadenylates and 2′5′ co-oligonucleotides

Cited by 64 publications

References 12 publications

Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Retinoic acid enhances the expression of interferon-induced proteins: evidence for multiple mechanisms of action

Methods for the Determination of the Interferon-Induced Enzyme 2'-5' Oligoadenylate Synthetase in Mononuclear Blood Cells

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