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Dolichyl phosphate linked sugars as intermediates in the synthesis of yeast mannoproteins: an in vivo study

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Abstract

Freshly prepared protoplasts of Saccharomyces cerevisiae X 2180 incorporate [3H]mannose and [14C]glucose for about 30 min into glycolipids and mannoproteins. Among the radioactive glycolipids formed dolichyl phosphate mannose, dolichyl phosphate glucose and dolichyl pyrophosphate oligosaccharides have been identified. The oligosaccharides released by weak acid from the dolichyl pyrophosphate were treated with endo-N-acetylglucosaminidase H and separated by gel filtration on Bio-Gel P-4. The largest oligosaccharide obtained corresponded exactly in size to Glc3Man9GlcNAc1 the compound formed also in animal tissues. Other oligosaccharides released from dolichyl pyrophosphate in addition to the glucose containing ones were mainly Man9GlcNAc1 and Man8GlcNAc1. No mannosyl oligosaccharide corresponding in size to the total “inner core” region found in native mannoproteins could be detected in a lipid-bound form.

The radioactive dolichyl pyrophosphate oligosaccharides were formed transiently; after 40 min only about 40% of the maximal radioactivity was observed in this fraction. In the presence of cycloheximide this decrease did not take place.

It is concluded that the dolichol pathway of N-glycosylation of glycoproteins in yeast cells is very similar, if not identical, to the reaction sequence worked out for animal cells.

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Authors and Affiliations

  1. Institut für Botanik, Fakultät für Biologie und Vorklinische Medizin, Universität Regensburg, D-8400, Regensburg, Federal Republic of Germany

    Ludwig Lehle, Ingrid Schulz & Widmar Tanner

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  1. Ludwig Lehle
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  2. Ingrid Schulz
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  3. Widmar Tanner
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Dedicated to Professor Dr. Otto Kandler on his 60th birthday

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Lehle, L., Schulz, I. & Tanner, W. Dolichyl phosphate linked sugars as intermediates in the synthesis of yeast mannoproteins: an in vivo study. Arch. Microbiol. 127, 231–237 (1980). https://doi.org/10.1007/BF00427198

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  • DOI: https://doi.org/10.1007/BF00427198

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