Browsing by Author "Pier Luigi Ipata"

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    Spectrophotometric Determination of Ribose-1 -Phosphate
    (1977) Umberto Mura; Francesco Sgarella; Pier Luigi Ipata
    In the course of studies on nucleoside monophosphate metabolism, the need was encountered for a method to determine ribose-1-phosphate. Published assays for ribose-1-phosphate depend either on chromatographic separation of the sugar-phosphate, or else on its acid lability which allows it to be determined as a phosphate. The present work describes a less laborious Spectrophotometric assay which is both rapid and specific. The basis of the method is the ab-sorbance change at 265 nm associated with the following two-stage enzymatic conversion: ribose- 1-phosphate + adenine = phosphate + adenosine (adenosine phosphorylase); adenosine + H2O —>inosine + NH3 (adenosine deaminase). The change in absorbance was proportional to ribose-1-phosphate concentration at least up to 25 /Ag/ml. In tests of the assay, it was possible to detect ribose-1-phosphate formation from inosine and phosphate catalyzed by purine nucleoside phosphorylase. Further, the degradation of ribose-1-phosphate by various commercial phosphatases and several tissues or microbial extracts was observed.