Iorio Egidio, Torosantucci Antonella, Bromuro Carla, Chiani Paola, Ferretti Amalia, Giannini Massimo, Cassone Antonio, Podo Franca
Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Viale Regina Elena, 299, Roma 00161, Italy.
Carbohydr Res. 2008 May 5;343(6):1050-61. doi: 10.1016/j.carres.2008.02.020. Epub 2008 Feb 29.
The structure of immunogenic and immunomodulatory cell wall glucans of Candida albicans is commonly interpreted in terms of a basic polysaccharide consisting of a beta-D-(1-->3)-linked glucopyranosyl backbone possessing beta-D-(1-->6)-linked side chains of varying distribution and length. This proposed molecular architecture has been re-evaluated by the present study on the products of selective enzymolysis of insoluble C. albicans glucan particles (GG). High resolution 1H (400 and 700 MHz) and 13C (100 and 175 MHz) NMR analyses were performed on a soluble beta-glucan preparation (GG-Zym) obtained by GG digestion with endo-beta-D-(1-->3)-glucanase and on its high- (Pool 1) and low-molecular weight (Pool 2) sub-fractions. The resonances typical of uniformly beta-D-(1-->6)- and beta-D-(1-->3)-linked linear glucans, together with additional multiplets assigned to short-chain oligoglucosides, were detected in GG-Zym. Pool 1 (46.3+/-6.4% of GG-Zym content) consisted of beta-D-(1-->6)-linked glucopyranosyl polymers, with short beta-D-(1-->3)-branched side chains of 2.20+/-0.02 units (branching degree (DB)=0.14+/-0.03). Pool 2 was a mixture of glucose and linear short-chain beta-D-(1-->3)-oligoglucosides. Further digestion of Pool 1 by beta-D-(1-->6)-glucanase yielded a mixture of glucose and short beta-D-(1-->6)-linked, either linear or beta-D-(1-->3,6) branched, oligomers. These endoglucanase digestion patterns were consistent with the presence in C. albicans cell wall glucans of beta-D-(1-->6)-linked glucopyranosyl backbones possessing beta-D-(1-->3)-linked side chains, a structure very close to that of beta-D-(1-->6)-glucan from Saccharomyces cerevisiae yeast. This finding may provide the grounds for further elucidation of the cell wall structure and a better understanding of the biological properties of C. albicans beta-glucans.
白色念珠菌具有免疫原性和免疫调节作用的细胞壁葡聚糖的结构,通常被解释为一种基本多糖,它由具有不同分布和长度的β-D-(1→3)连接的吡喃葡萄糖基主链以及β-D-(1→6)连接的侧链组成。本研究通过对不溶性白色念珠菌葡聚糖颗粒(GG)的选择性酶解产物,对这一提出的分子结构进行了重新评估。对通过用内切β-D-(1→3)-葡聚糖酶消化GG得到的可溶性β-葡聚糖制剂(GG-Zym)及其高分子量(组分1)和低分子量(组分2)亚组分进行了高分辨率1H(400和700 MHz)和13C(100和175 MHz)NMR分析。在GG-Zym中检测到了均匀的β-D-(1→6)-和β-D-(1→3)-连接的线性葡聚糖的典型共振,以及归属于短链低聚葡糖苷的额外多重峰。组分1(占GG-Zym含量的46.3±6.4%)由β-D-(1→6)-连接的吡喃葡萄糖基聚合物组成,带有2.20±0.02个单位的短β-D-(1→3)-分支侧链(分支度(DB)=0.14±0.03)。组分2是葡萄糖和线性短链β-D-(1→3)-低聚葡糖苷的混合物。用β-D-(1→6)-葡聚糖酶对组分1进行进一步消化,得到了葡萄糖和短的β-D-(1→6)-连接的(线性或β-D-(1→3,6)分支的)低聚物的混合物。这些内切葡聚糖酶的消化模式与白色念珠菌细胞壁葡聚糖中存在具有β-D-(1→3)-连接侧链的β-D-(1→6)-连接的吡喃葡萄糖基主链一致,这一结构与酿酒酵母的β-D-(1→6)-葡聚糖非常接近。这一发现可能为进一步阐明细胞壁结构以及更好地理解白色念珠菌β-葡聚糖的生物学特性提供依据。