Identification of Inositol 1,3,4-Trisphosphate 5-Kinase and Inositol 1,3,4,5-Tetrakisphosphate 6-Kinase in Immature Soybean Seeds

In extracts of immature soybean (Glycine max [L.] Merr.) seeds inositol tetrakisphosphate was formed from [3H]inositol 1,3,4-trisphosphate but not from [3H]inositol 1,4,5-trisphosphate. Inositol 1,3,4-trisphosphate kinase was purified to a specific activity of 3.55 min−1 mg−1 by polyethylenimine clarification and anion-exchange chromatography. The partially purified enzyme converted [3H]inositol 1,3,4-trisphosphate to inositol 1,3,4,5-tetrakisphosphate as the major product and inositol 1,3,4,6- and/or 1,2,3,4-tetrakisphosphate as the minor product. Subsequent experiments revealed a separate inositol 1,3,4,5-tetrakisphosphate 6-kinase activity, which could link these enzymes to inositol hexakisphosphate synthesis via the previously reported inositol 1,3,4,5,6-pentakisphosphate 2-kinase. The apparent Km values for inositol 1,3,4-trisphosphate kinase were 200 ± 0 nm for inositol 1,3,4-trisphosphate and 171 ± 4 μm for ATP, and the reaction was not reversible. The kinetics were such that no activity could be detected using unlabeled inositol 1,3,4-trisphosphate and [γ-32P]ATP, which suggested that other kinases may have been observed when less purified fractions were incubated with radiolabeled ATP. Inositol 1,3,4-trisphosphate kinase was nonspecifically inhibited more than 80% by various inositol polyphosphates at a concentration of 100 μm.

(Table S1) Radiocarbon data of sediment core SO161/5 22SL

At the end of the Last Glacial Maximum (19,000 to 11,000 years ago), atmospheric carbon dioxide concentrations rose while the Delta14C of atmospheric carbon dioxide declined**1, 2. These changes have been attributed to an injection of carbon dioxide with low radiocarbon activity from an oceanic abyssal reservoir that was isolated from the atmosphere for several thousand years before deglaciation**3. The current understanding points to the Southern Ocean as the main area of exchange between these reservoirs4. Intermediate water formed in the Southern Ocean surrounding Antarctica would have then carried the old carbon dioxide signature to the lower-latitude oceans**5, 6. Here we reconstruct the Delta14C signature of Antarctic Intermediate Water off the coast of Chile for the past 20,000 years, using paired 14C ages of benthic and planktonic foraminifera. In contrast to the above scenario, we find that the delta14C signature of the Antarctic Intermediate Water closely matches the modelled surface ocean Delta14C, precluding the influence of an old carbon source. We suggest that if the abyssal ocean is indeed the source of the radiocarbon-depleted carbon dioxide, an alternative path for the mixing and propagation of its carbon dioxide may be required to explain the observed changes in atmospheric carbon dioxide concentration and radiocarbon activity.