Biosynthesis of purines by a proplastid fraction from soybean nodules

Abstract

A proplastid-containing fraction was rapidly prepared from soybean nodules by a combination of differential and step gradient centrifugation. This fraction was capable of incorporating [U-14C]glycine into purines in the presence of added phosphoribosylpyrophosphate or ribose 5-phosphate, glutamine, aspartate, ATP, bicarbonate, methenyl tetrahydrofolate, MgCl2, and KCl. The primary product was IMP; some inosine was also formed. Soluble and bacteroid fractions from soybean nodules gave considerably lower rates of incorporation. Labeled carbon from both [U-14C]serine and [3-14C]serine was incorporated into purines when tetrahydrofolate and NADP+ were substituted for methenyl tetrahydrofolate. In this case, small amounts of label were also found in AMP and xanthine monophosphate (XMP). Labeled bicarbonate was incorporated into IMP and inosine by the proplastid fraction. Labeled formate, however, was not a competent substrate for purine synthesis, indicating the absence of formyl tetrahydrofolate synthetase activity in this fraction. When labeled IMP was incubated with a proplastid preparation, most of the label appeared in inosine. XMP and xanthosine were also formed if NAD+ or NADP+ was added to the incubation mixture indicating the presence of IMP dehydrogenase activity in the proplastid fraction.