Sphingomyelin synthesis was studied in slices of rat heart by using [Me-14C]choline, [1,2-14C]ethanolamine, S-adenosyl-L-[14C]methionine and [32P]Pi as as precursors. In the presence of both [Me-14C]choline and [32P]Pi the ratio of the specific radioactivities of 14C and 32P in phosphatidylcholine was greater than in sphingomyelin at all the times studied. This suggested that synthesis of phosp...

“Light” nerve endings from guinea pig cerebral cortex were incubated with [32P]orthophosphate, myo[3H]inositol, [3H]glycerol, or [3H]glucose in the presence or absence of cholinergic agents. Acetylcholine and carbamylcholine increased incorporation of “Pi into phosphatidic acid 100% and into various classes of phosphatidylinositol 20 to 70%. These agents also stimulated the labeling of phosphat...

A recombinant protein-tyrosine-phosphatase has been expressed in Escherichia coli and purified to a single band by sodium dodecyl sulfate-polyacrylamide gel electrophoresis using affinity chromatography. When the phosphatase was allowed to react with 32Plabeled substrates and then rapidly denaturated, a 32Plabeled phosphoprotein could be visualized by sodium dodecyl sulfate-polyacrylamide gel ...

The 3' terminus of the strand (minus strand) complementary to poliovirion RNA (plus strand) has been examined to see whether this sequence extends to the 5'-nucleotide terminus of the plus strand, or whether minus-strand synthesis terminates prematurely, perhaps due to the presence of a nonreplicated nucleotide primer for initiation of plus-strand synthesis. The 3' terminus was labeled with 32P...

The phospholipids of cultured parsley and soybean cells were labelled with myo-[2-3H]inositol, [2-3H]glycerol or [32P]orthophosphate. By oneand two-dimensional chromatographic compari­ son of the labelled phospholipids with reference substances, the presence of l-(3-sn-phosphatidyl)-D-myo-inositol 4-phosphate and l-(3-sn-phosphatidyl)-D-myo-inositol 4,5-bisphosphate was demonstrated in these cu...

Hansenula polymorpha Pex14p (HpPex14p) is a component of the peroxisomal membrane essential for peroxisome biogenesis. Here, we show that HpPex14p is phosphorylated in vivo. In wild-type H. polymorpha cells, grown in the presence of [32P]orthophosphate, the 32P label was incorporated into HpPex14p. Labelled HpPex14p was induced after a shift of cells to methanol-containing media and rapidly dis...

The sensitivity, rapidity, and ease of use of biotinylated (Blugene, Bethesda Research Laboratories) and 32P cDNA probes have been compared, the probe being the cDNA for beta-glucocerebrosidase (EC 3.1.2.45). With the Blugene kit I could detect 2 pg of biotinylated DNA on dot blots. However, under conditions of hybridization, the lower limit of detection for unlabeled cDNA (transblotted onto ni...

We describe an enzymic quantification of total hydrolyzable glycerol in amniotic fluid. An extract of amniotic fluid is hydrolyzed in mild alkali, without pretreatment with lipase. The resulting glycerol (some of which may be derived from trace amounts of triglyceride) is then enzymatically phosphorylated with adenosine 5'-[gamma 32P]triphosphate to yield glycerol [gamma 32P]phosphate. After re...