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A glucanase-driven fractionation allows redefinition of Schizosaccharomyces pombe cell wall composition and structure: assignment of diglucan.
Anal Biochem. 2005 Jan 15; 336(2):202-12.AB

Abstract

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.

Authors+Show Affiliations

Magnelli PE
Department of Molecular and Cell Biology, School of Dental Medicine, Boston University, 15 Stoughton St., EVANS 4, Boston, MA 02118, USA.
Cipollo JF
No affiliation info available
Robbins PW
No affiliation info available

MeSH

Carbohydrate SequenceCell WallDialysisEndo-1,3(4)-beta-GlucanaseGlucansMagnetic Resonance SpectroscopySchizosaccharomyces

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15620885

Citation

Magnelli, Paula E., et al. "A Glucanase-driven Fractionation Allows Redefinition of Schizosaccharomyces Pombe Cell Wall Composition and Structure: Assignment of Diglucan." Analytical Biochemistry, vol. 336, no. 2, 2005, pp. 202-12.
Magnelli PE, Cipollo JF, Robbins PW. A glucanase-driven fractionation allows redefinition of Schizosaccharomyces pombe cell wall composition and structure: assignment of diglucan. Anal Biochem. 2005;336(2):202-12.
Magnelli, P. E., Cipollo, J. F., & Robbins, P. W. (2005). A glucanase-driven fractionation allows redefinition of Schizosaccharomyces pombe cell wall composition and structure: assignment of diglucan. Analytical Biochemistry, 336(2), 202-12.
Magnelli PE, Cipollo JF, Robbins PW. A Glucanase-driven Fractionation Allows Redefinition of Schizosaccharomyces Pombe Cell Wall Composition and Structure: Assignment of Diglucan. Anal Biochem. 2005 Jan 15;336(2):202-12. PubMed PMID: 15620885.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A glucanase-driven fractionation allows redefinition of Schizosaccharomyces pombe cell wall composition and structure: assignment of diglucan. AU - Magnelli,Paula E, AU - Cipollo,John F, AU - Robbins,Phillips W, PY - 2004/06/21/received PY - 2004/12/29/pubmed PY - 2005/5/5/medline PY - 2004/12/29/entrez SP - 202 EP - 12 JF - Analytical biochemistry JO - Anal Biochem VL - 336 IS - 2 N2 - 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. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/15620885/A_glucanase_driven_fractionation_allows_redefinition_of_Schizosaccharomyces_pombe_cell_wall_composition_and_structure:_assignment_of_diglucan_ DB - PRIME DP - Unbound Medicine ER -