Purification of an autocatalytic protein-glycosylating enzyme from cell suspensions of Daucus carota L.
A glycosyltransferase was identified in the 174 000 · g membrane pellet and the supernatant from extracts of cell suspensions of Daucus carota L. The enzyme from the supernatant was enriched 475-fold, and sodium dodecyl sulfate-gel electrophoresis and fluorography of this purified sample showed that the only enriched protein band (40 000 Da) was simultaneously an enzyme and a glucose-acceptor. Gel filtration and electrophoresis under non-denaturing conditions proved that in vivo this protein provides the subunits for a very large molecule. Radio-gas-liquid chromatography demonstrated that only one glucosyl moiety was transferred from UDP-glucose to the protein.
Auf der Morgenstelle 1, Institut für Biologie I der Universität, D-7400, Tübingen, Federal Republic of Germany.
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Quentmeier
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Ingold
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Seitz