Journal
ANALYTICAL BIOCHEMISTRY
Volume 336, Issue 2, Pages 202-212Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2004.09.022
Keywords
diglucan; beta-glucan; alpha 1,3-glucanase; cell wall; Schizosaccharomyces; fungi
Funding
- NCRR NIH HHS [S10 RR15942, P41 RR10888] Funding Source: Medline
- NIGMS NIH HHS [GM 31318] Funding Source: Medline
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Purified endoglucanases have been used to determine the composition of Schizosaccharomyces pombe cell wall. This structure has been traditionally studied after isolating its components (mannoproteins, alpha1,3-glucan, beta1,3-glucan, and a branched beta-glucan) with hot alkali. Instead, we sequentially removed the polysaccharides by digesting with endo-beta1,3-glucanase and with a novel endo-alpha1,3-glucanase (mutanase). After this gentle isolation we observed that a branched beta1,3-beta1,6-glucan is much more abundant than previously described. By scaling-up the new protocol we prepared large amounts of the highly branched glucan and determined its structural features. We have named this highly branched beta-glucan diglucan, reflecting its two types of beta linkages. We have also identified an insoluble endoglucanase-resistant type of 1,3-linked glucan present in S. pombe cell walls. We redefined the wall composition of S. pombe vegetative cells by this new method. Finally, to demonstrate its application, we determined the cell wall composition of known mutant strains. (C) 2004 Published by Elsevier Inc.
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