Spectrophotometric determination of ribose-1-phosphate

✍ Scribed by Umberto Mura; Francesco Sgarrella; Pier Luigi Ipata

• Publisher: Elsevier Science
• Year: 1977
• Tongue: English
• Weight: 407 KB
• Volume: 82
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(77)90150-6

✦ Synopsis

In the course of studies on nucleoside monophosphate metabolism, the need was encountered for a method to determine ribose-l-phosphate. Published assays for ribose-l-phosphate depend either on chromatographic separation of the sugarphosphate, 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 absorbance change at 265 nm associated with the following two-stage enzymatic conversion: ribose-l-phosphate + adenine ti phosphate + adenosine (adenosine phosphorylase); adenosine + HZ0 + inosine + NH, (adenosine deaminase). The change in absorbance was proportional to ribose-l-phosphate concentration at least up to 25 CLg/ml. In tests of the assay, it was possible to detect ribose-lphosphate formation from inosine and phosphate catalyzed by purine nucleoside phosphorylase. Further, the degradation of ribose-l-phosphate by various commercial phosphatases and several tissues or microbial extracts was observed.