Regeneration of Candida albicans protoplasts began with the formation of a chitin network which was complemented after a lag of about 60 min by the deposition of beta-glucan. Proteins were incorporated early to the growing structure, beginning with the mannoproteins which are kept in place by non-covalent bonds. Incorporation of covalently linked mannoproteins took place only after deposition of glucan. The incorporation of these mannoproteins did not occur when protoplasts were incubated with papulacandin B which inhibited glucan formation, or with tunicamycin which blocked N-glycosylation of mannoproteins. In the presence of papulacandin B, large amounts of native mannoproteins accumulated in the medium. However, in the presence of tunicamycin, the large mannoprotein material found was of smaller apparent molecular weight, suggesting that it was deficient in glycosylation. Partially regenerated walls were able to incorporate 'in vitro' non-covalently bound mannoproteins, indicating that some components of very large cellular structures such as walls are capable of being articulated by a self-assembly process.
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