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A fluorescent substrate for the continuous assay of phosphatidylinositol-specific phospholipase C: synthesis and application of 2-naphthyl myo-inositol-1-phosphate


, : A fluorescent substrate for the continuous assay of phosphatidylinositol-specific phospholipase C: synthesis and application of 2-naphthyl myo-inositol-1-phosphate. Analytical Biochemistry 198(1): 10-14

A fluorescent water-soluble substrate for phosphatidylinositol-specific phospholipase C was synthesized. The diacylglycerol moiety of the natural substrate, phosphatidylinositol, was replaced by the fluorescent moiety, 2-naphthol, resulting in the synthetic substrate, racemic 2-naphthyl myo-inositol-1-phosphate. The synthetic substrate provided a continuous fluorometric assay for the phosphatidylinositol-specific phospholipase C from Bacillus cereus. Initial rates of the cleavage of the 2-naphthyl substrate by the phospholipase measured by fluorometry were linear with time and the amount of enzyme added. The specific enzyme activity at pH 8.5 and 25 degrees C was about 0.04 mumol/min mg protein at an initial substrate concentration of 0.8 mM. 31P NMR experiments suggest that, as with phosphatidylinositol itself, cleavage of the fluorescent substrate proceeds in two steps via a myo-inositol-1,2-cyclic phosphate intermediate, and that only the D-isomer is a substrate for the B. cereus phospholipase. The synthetic substrate was stable during long-term storage as a solid in the dark at -20 degrees C. It was also stable for several weeks when stored in the dark frozen in aqueous solution near neutral pH.

US$29.90

PMID: 1789410


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Tanaka, Shigeho, 1931: On the distribution of fishes in Japanese waters. This paper is restricted to the fish fauna of Hokkaido, Hondo, Shikoku, and Kiushiu. The author first discusses a number of examples of variation according to individual, age, sex, season, and locality. Colored plates show 3 color varieties of Bra...

Harms, K.J.; Tovar, K.R.; Craig, A.Marie., 2005: Synapse-specific regulation of AMPA receptor subunit composition by activity. We examined the changes that arise when neurotransmitter release is inhibited in a subpopulation of hippocampal neurons in coculture with normally active neighbors. Subsets of neurons were presynaptically silenced by chronic expression of tetanus...

Langhe, E.D.; Vermeulen, J., 1972: Evolution of abscisic acid in the tissues of a cotton boll. Levels of ABA during maturation of cotton bolls were monitored in the pericarp, fibre, testa and embryo + endosperm. The embryo + endosperm contained high levels of ABA 21-25 days after anthesis. The increase in ABA at 35-50 days after anthesis wa...

Chang C.S., 1986: Nitrogen oxides control techniques suitable to developing countries. Atmospheric Environment 20(10): 2071

Teumer B., 1976: Electro myographic documentation and automatic report generation. Electroencephalography & Clinical Neurophysiology: 668-669

Noss, G., 1987: Human polyoma virus type BK infection and T antibody response in renal transplant recipients. BKV infections were detected in 103 of 168 renal transplant recipients. 18 patients showed primary infection within 3 months after transplantation as evidenced by seroconversion and virus isolation. Secondary BKV infection occurred in 44 patients...

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Romans, William W., 1999: Belted seed metering device. An seed metering device and method for disbursing agricultural seed and the like singularly and accurately comprising sprocket wheels, a seed conveyor device, a housing. A seed conveyor device interlocks with and surrounds sprocket wheels and rota...

Dahl, G.M.; von Wolcker, E., 1968: On possibilities and planning of regional installations for water fluoridation. Tidning. Sveriges Tandlakarforbund 60(17): 774-779