Radioimmune, radiobinding and HPLC analysis of 2-5A and related oligonucleotides from intact cells
- 13 November 1980
- journal article
- research article
- Published by Springer Nature in Nature
- Vol. 288 (5787) , 189-192
- https://doi.org/10.1038/288189a0
Abstract
The enzyme (2-5 A synthetase) which synthesizes ppp(A2′p)n A where n = 2 to 4 (collectively referred to as 2-5A)1 is widely distributed in a variety of cells and tissues in amounts which increase in response to interferon2–5 and vary with growth and hormone status6. 2-5A activates a nuclease which inhibits protein synthesis7–10. The non-phosphorylated ‘core’ of 2-5A ((A2′p)nA, n = 2 to 4) can inhibit DNA synthesis and cell growth11,12. Here we describe convenient and sensitive radioimmune (RI) and radiobinding (RB) assays for core and 2-5A. In combination with more satisfactory high performance liquid chromatography (HPLC) methods using reverse-phase C18 columns, these assays have been used to detect core and 2-5A in crude extracts from interferon-treated cells. The novel 2-5A synthetase products NAD2′p5′A2′p5′A and A5′p45′A2′p5′A2′p5′A (ref. 13), which can also be detected using the RB assay, were not found in significant amounts. The natural occurrence of core has not been described previously.Keywords
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