Purification and properties of guanine, queuine-tRNA transglycosylase from wheat germ.

Abstract

Guanine, queuine-tRNA transglycosylase has been purified from wheat germ to homogeneity. The specific activity is 2,000 pmol h-1 mg-1 of protein. The enzyme has an apparent Mr = 140,000. It migrates as a single band with Mr = 68,000 on electrophoresis in sodium dodecyl sulfate gels, indicating two Mr = 68,000 subunits. Both guanine (Km = 6.0 X 10(-8) M) and queuine (KI = 9.5 X 10(-8) M) are substrates but 7-(aminomethyl)-7-deazaguanine is not. The enzyme requires a monovalent or a divalent cation; Na+ and Mg2+ are more effective activators than other cations. The optimum pH is 7.6. Six tRNA isoacceptors found in wheat germ are substrates for the enzyme.