A spectrophotometric method for determination of sphingomyelinase

S. Gatt*, T. Dinur, Y. Barenholz

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

A colored derivative of sphingomyelin was synthesized and used as substrate for several sphingomyelinases. The compound is N-ω-trinitrophenyl aminolaurylsphingosylphosphorylcholine. The rate of hydrolysis of this substrate was compared to that of bovine brain sphingomyelin, labelled with tritium in the choline moiety. The following enzyme preparations were used: homogenate-less debris of brain, assayed at pH 5.0 or 7.4; a solubilized preparation derived from rat brain lysosomes, assayed at pH 5.0 and a purified enzyme of Staphylococcus aureus. With all preparations, the rates of hydrolysis of the yellow derivative were very similar to those of the brain sphingomyelin. Extracts of skin fibroblasts of normal and Niemann-Pick patients as well as amniotic cells were also used. Again, the rates of hydrolysis of the yellow derivative practically equalled those using brain sphingomyelin.

Original languageEnglish
Pages (from-to)503-507
Number of pages5
JournalBiochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Volume530
Issue number3
DOIs
StatePublished - 28 Sep 1978

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