Synthesis of hyaluronate in differentiated teratocarcinoma cells. Characterization of the synthase

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Abstract
Differentiation of teratocarcinoma cells [F9] led to induction of hyaluronate synthesis. The synthase was recovered in the membrane fraction of cell lysates. Hyaluronate was synthesized at the membranes and was then released as a soluble product. The synthase could be stimulated by a variety of phosphate esters which prevented the degradation of the substrates UDP-GlcNAc [N-acetyl glucosamine] and UDP-GlcA [glucosamine] and the release of the growing hyaluronic acid chain from the membrane. Hyaluronidases or oligosaccharides derived from hyaluronate did not affect the synthesis. The chains grew at a rate of 60 repeating U/min. Continuous new chain initiation occurred during prolonged synthesis. Digestion of pulse-chase-labeled hyaluronate with .beta.-N-acetylglucosaminidase and .beta.-glucuronidase showed that the chains grew at the reducing end.