Glucan Synthesis in Pneumocystis carinii |
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Authors: | DEBRA J. WILLIAMS JEFFREY A. RADDING ANNE DELL KAY-HOOI KHOO MARK E. ROGERS FRANK F. RICHARDS MARTINE Y. K. ARMSTRONG |
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Affiliation: | MacArthur Center for Molecular Parasitology, Yale University School of Medicine, 60 College Street, New Haven, Connecticut 06510, USA;Department of Biochemistry, Imperial College of Science, Technology and Medicine, Exhibition Road, London S.W.7, United Kingdom;M-SCAN Ltd., Silwood Park, Berkshire SL5 7PZ, United Kingdom |
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Abstract: | Rat-derived Pneumocystis carinii lysed with sodium deoxycholate catalysed the incorporation of uridine diphospho-glucose into an insoluble polymer. This enzyme activity was present in both the pellet and the supernatant when the P. carinii preparations were centrifuged. The polymer whose production was catalysed by the supernatant was examined by mass spectrometry and found to be an α 1→4 glucan, which is either unbranched or has relatively few branches. Polymer formation was completely inhibited by the addition of α amyloglucohydrolase to the supernatant. Polymer formation in the pellet of deoxycholate P. carinii preparations, unlike that in the supernatant, was partially resistant to α amyloglucohydrolase. The soluble glucan synthase activity in the supernatant was stable for more than 30 h at room temperature and was approximately 50 times more active on a cell-to-cell basis than the supernatant from deoxycholate preparations of the yeast Saccharomyces cerevisae. |
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Keywords: | αL 1-4 glucan cell culture mass spectrometry rat uridine diphosphoglucose. |
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