Properties of a Protein Activator of NAD Kinase from Plants

Abstract

Purification of pea (Pisum sativum) seedling NAD kinase by DEAE cellulose column chromatography resulted in loss of activity, due to dissociation of an activator from the enzyme. The purified enzyme preparation, which was almost completely inactive, regained the activity when the activator was added back. The activator was purified 320-fold by ion exchange chromatographies. The activator was susceptible to proteolytic enzymes, but not to ribonuclease, glucoamylase or pectinase, indicating that it is of a protein nature. This protein was relatively stable in boiling water, but susceptible to acid or alkali, especially under high temperatures. Restoration of catalytic activity of inactive enzyme was proportional to amounts of the activator added. Gel filtration indicated that MW of the activator was 28,000. The activator was found in extracts from various plants [Zea mays, Oryza sativa, Chlorella vulgaris, Spinacia oleracea, Brassica rapa and Pisum sativum].