Cytidylate cyclase: The product isolated by the method of Cech and Ignarro is not cytidine 3′,5′-monophosphate

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Abstract

When mouse liver homogenates were incubated with [α-32P]-CTP according to the method of Cech and Ignarro (4,5), a [32P] product (100–120 pmoles/mg protein/minute) was isolated by chromatography on alumina column, in the same fraction as [3H]-cyclic CMP.1 However this product did not behave as cyclic CMP in various chromatographic systems. Moreover the amount of this [32P] product isolated was markedly reduced by removal of protein prior to chromatography on alumina. Chromatography of the enzymatic reaction product either before or after alumina purification on Dowex 1 — formate column indicated that 5′-CMP and CDP are the major products, with no formation of cyclic CMP. These results indicate that the [32P] labeled material isolated by Cech and Ignarro is mainly if not entirely 5′-CMP and CDP.

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Abbreviations: Cyclic CMP, cytidine 3′,5′-monophosphate.

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