Metabolic regulation during early frog development. Identification of proteins labeled by 32P-glycolytic intermediates

• Published in: The Journal of biological chemistry 1987-12, Vol.262 (35), p.17038-17045
• Main Authors: Dworkin, M B, Dworkin-Rastl, E
• Format: Article
• Language: English
• Subjects: 2,3-Diphosphoglycerate; Adenosine Triphosphate - metabolism; Animals; Biological and medical sciences; Cell metabolism, cell oxidation; Cell physiology; Diphosphoglyceric Acids - metabolism; Fundamental and applied biological sciences. Psychology; Gluconeogenesis; Glycogen - metabolism; Glycolysis; Molecular and cellular biology; Molecular Weight; Phosphoenolpyruvate - metabolism; Phosphoglycerate Mutase - metabolism; Proteins - analysis; Sodium Fluoride - pharmacology; Xenopus - embryology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)45488-9

When 32P-labeled phosphoenolpyruvate is injected into Xenopus laevis oocytes, a 50-60-kDa protein of subunit size Mr 29,000 is rapidly labeled, followed by a second (monomeric) protein of 66 kDa concomitant with the loss of label from the first protein. We have identified these proteins as, respectively, the glycolytic enzymes phosphoglyceromutase and phosphoglucomutase. The phosphoglyceromutase is labeled at a histidine and the phosphoglucomutase at a serine, presumably at their active sites during the gluconeogenic transformation of phosphoenolpyruvate into glycogen. The transfer of the 32P label from phosphoenolpyruvate to these two enzymes also occurs in in vitro lysates made from full-grown Xenopus oocytes, eggs, or early embryos, but with a slower time course. Lysates prepared from leg muscle show labeling of the phosphoglyceromutase, but not the phosphoglucomutase, when incubated with [32P]phosphoenolpyruvate. This last result is expected in tissues showing metabolic flux largely in the glycolytic direction. The data indicate that in full-grown oocytes and embryos metabolic flux occurs largely in the gluconeogenic direction.