Cyst wall synthase: -acetylgalactosaminyltransferase activity is induced to form the novel -acetylgalactosamine polysaccharide in the cyst wall Free

Abstract

Uridine-5′-diphospho--acetylgalactosamine (UDP-GalNAc) is required in the formation of the outer filamentous wall of and is synthesized by inducible enzymes in the cytosol of encysting trophozoites. In this study, an inducible enzyme activity that is associated with a particle population isolated from encysting is reported, and this activity exclusively incorporates [1-C]GalNAc (from UDP-[C]GalNAc) into an ethanol precipitate with the same properties as the filamentous cyst wall of . This ethanol precipitate exhibits characteristics of cyst wall filaments in that both contain GalNAc as the only sugar moieties and are SDS-insoluble, proteinase- and alkali-resistant and acid-hydrolysable. However, since the precise chemical nature of the ethanol precipitate remains unknown, this enzyme activity is referred to tentatively as cyst wall synthase (CWS). CWS activity peaks in cells between 24 and 36 h of encystment and exhibits a high affinity and marked specificity for UDP-GalNAc as its substrate. UDP--acetylglucosamine, UDP-glucose, UDP-galactose, -glucosamine and -galactosamine were not incorporated into the ethanol precipitate. Partially purified CWS activity exhibits an apparent of 0·048 mM for UDP-GalNAc, a of 0·70 nmol min (mg protein) and a requirement for divalent cations in the following order of preference: Ca, Mg>Co>>>Mn, Zn. EDTA inhibits CWS activity.

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2004-05-01
2024-03-28
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