Determination of Specific Radioactivities of Mononucleotides and Glycolytic Intermediates from Mouse Liver after Labelling in vivo with [32P]Orthophosphate

• DOI: 10.1111/j.1432-1033.1968.tb00458.x
• Publication Date: 2005-03-03T12:25:47Z
• Authors: U. Till; E. Blume; J. Günther; J.G. Reich; D. Zahn; R. Klinger; K. Jaroszewicz; H. Frunder
• Keywords: Dihydroxyacetone phosphate; Dihydroxyacetone; Specific activity

Experimental procedures and conditions are described for the determination of specific radioactivities 16 different P ‐positions in 12 phosphorylated liver compounds after labelling vivo with [ 32 P]orthophosphate. These data necessary calculations flux rates within mononucleotides glycolytic pathway. Purification following given: ATP, ADP, GTP, UTP, UDPG, glucose 6‐phosphate, fructose 1,6‐biphosphate, dihydroxyacetone‐phosphate, glycerol 3‐phosphate, phosphoglyceric acid phosphoenolpyruvate. The purification each compound consists 3–5 chromatographic steps, most cases including a enzymic conversion. A preparation procedure is mouse which shows only negligible artificial isotope exchange between β‐ γ‐ ATP. This was proved by addition labelled ATP to non‐labelled livers. Five ten min intravenous injection carrier‐free P]orthophosphate examined show identical values, thereby excluding compartmentations kinetic relevance.