Abstract
Publisher Summary This chapter discusses the synthesis of colominic acid synthetase. Colominic acid is a homopolymer of one of the sialic acids, N -acetylneuraminic acid (NAN). The extent of incorporation of sialic acid into colominic acid is determined using CMP-1- 14 C-NAN (cytidine 5′-monophospho- N -acetylneuraminic acid) as substrate. The substrate and its degradation product, NAN, are separated by paper chromatography from the polymer, which remains at the origin. CMP-NAN can also be separated from colominic acid, which remains near the origin, by electrophoresis on Whatman No. 3 MM paper for 30 minutes at 90 volts/cm in 1% sodium tetraborate, pH 9.0. The ammonium sulfate in the incubation mixture interferes with the resolution, but the procedure is useful for a quick survey of activity. The reagents used, procedure followed, and the steps involved in the purification are also described in the chapter. CMP-NAN is the immediate donor of NAN for incorporation into colominic acid.
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