Characterisation of Purine Nucleoside Phosphorylase from Fibroblasts Using Ultra-Microchemical Methods

Abstract

A new technique to quantitate nucleoside phosphorylase (NP) activity in single or small numbers of counted visually selected cells is presented. Fibroblasts were cultivated on the plastic film bottom of culture dishes. After lyophilisation in situ, plastic film leaflets carrying a counted number of cells were cut out and tested for NP activity. Some properties of NP, including temperature stability, pH optimum and substrate affinity, have been studied. The data obtained suggest that NP might play a regulatory role in the purine interconversion pathway.