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(1
6)-GLUCAN INTERCONNECTS MANNOPROTEIN, (13)-GLUCAN,
AND CHITIN
(Received for publication, April 3, 1997)
,
,
,
,
and
From the In a previous study (Kollár, R.,
Petráková, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170-1178), the linkage
region between chitin and 
Laboratory of Biochemistry and Metabolism,
the 
Laboratory of Bioorganic Chemistry, and the ** Laboratory of
Cell Biochemistry and Biology, NIDDK, Bethesda, Maryland 20892, the
§ Mass Spectrometry Resource, Boston University School of
Medicine, Boston, Massachusetts 02118, the ¶ Center for Biologics
Evaluation and Research, Food and Drug Administration, Bethesda,
Maryland 20892, and the Institute of
Molecular Cell Biology, University of Amsterdam, 1098 SM
Amsterdam, The Netherlands
(13)-glucan was solubilized and
isolated in the form of oligosaccharides, after digestion of yeast cell
walls with (13)-glucanase, reduction with borotritide, and
subsequent incubation with chitinase. In addition to the
oligosaccharides, the solubilized fraction contained tritium-labeled
high molecular weight material. We have now investigated the nature of
this material and found that it represents areas in which all four
structural components of the cell wall, (13)-glucan,
(16)-glucan, chitin, and mannoprotein are linked together.
Mannoprotein, with a protein moiety about 100 kDa in apparent size, is
attached to (16)-glucan through a remnant of a
glycosylphosphatidylinositol anchor containing five 
-linked mannosyl
residues. The (16)-glucan has some (13)-linked branches,
and it is to these branches that the reducing terminus of chitin chains
appears to be attached in a (14) or (12) linkage. Finally,
the reducing end of (16)-glucan is connected to the nonreducing
terminal glucose of (13)-glucan through a linkage that remains to
be established. A fraction of the isolated material has three of the
main components but lacks mannoprotein. From these results and previous
findings on the linkage between mannoproteins and (16)-glucan, it
is concluded that the latter polysaccharide has a central role in the
organization of the yeast cell wall. The possible mechanism of
synthesis and physiological significance of the cross-links is
discussed.
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