A pyrophosphate bridge links the pyruvate-containing secondary cell wall polymer of Paenibacillus alvei CCM 2051 to muramic acid
A pyrophosphate bridge links the pyruvate-containing secondary cell wall polymer of Paenibacillus alvei CCM 2051 to muramic acid
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DOI：
10.1023/a:1011062302889
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发表时间：
2000-10-01
影响因子：
3
通讯作者：
Messner, P
中科院分区：
文献类型：
--
作者：
Schäffer, C;Müller, N;Messner, P
肽聚糖、次级细胞壁聚合物（SCWP）和表面层（S-层）糖蛋白是蜂房类芽孢杆菌CCM 2051的主要糖基化细胞壁组分。在本报告中，SCWP的完整结构，其连接的肽聚糖层，其物理化学性质进行了研究。根据化学和结构分析以及一维和二维核磁共振光谱的综合证据，提出了SCWP-肽聚糖复合物的以下结构：[（Pyr 4，6）-β-D-ManpNAc-（1 ->4）-β-D-GlcpNAc-（1 ->3）]（n类似于11）-（Pyr 4，6）-β-D-ManpNAc-（1 ->4）-α-D-GlcpNAc-（1 ->O）-PO 2-O-PO 2-（O ->6）-MurNAc-每个二糖单元被4，6-连接的葡糖酸残基取代。在温和的酸性条件下，高达50%的ManNAc会丢失，留下未取代的ManNAc残基。构成SCWP的阴离子聚糖链通过焦磷酸基团随机连接到肽聚糖层的胞壁酸残基的C-6。P-31 NMR揭示了两个信号，作为胶束形成的结果，经历不同的谱线增宽。因此，它们的积分比明显偏离1：1。通过用乙二胺四乙酸、十二烷基硫酸钠和在NMR测量之前立即进行超声处理，该比率接近1。这种行为的可逆性证实了存在焦磷酸连接在这个SCWP-肽聚糖complex.除了确定的SCWP的结构和连接，其生物学功能的一种可能的方案进行了讨论。
The peptidoglycan, the secondary cell wall polymer (SCWP), and the surface layer (S-layer) glycoprotein are the major glycosylated cell wall components of Paenibacillus alvei CCM 2051. In this report, the complete structure of the SCWP, its linkage to the peptidoglycan layer, and its physicochemical properties have been investigated. From the combined evidence of chemical and structural analyses together with one- and two-dimensional nuclear magnetic resonance spectroscopy, the following structure of the SCWP-peptidoglycan complex is proposed:[(Pyr4,6)-beta -D-ManpNAc-(1 -->4)-beta -D-GlcpNAc-(1 -->3)](n similar to 11)-(Pyr4,6)-beta -D-ManpNAc-(1 -->4)-alpha -D-GlcpNAc-(1 -->O)-PO2-O-PO2-(O -->6)-MurNAc-Each disaccharide unit is substituted by 4,6-linked pyruvic acid residues. Under mild acidic conditions, up to 50% of them are lost, leaving non-substituted ManNAc residues. The anionic glycan chains constituting the SCWP are randomly linked via pyrophosphate groups to C-6 of muramic acid residues of the peptidoglycan layer. P-31 NMR reveals two signals that, as a consequence of micelle formation, experience different line broadening. Therefore, their integral ratio deviates significantly from 1:1. By treatment with ethylenediaminetetraacetic acid, sodium dodecyl sulfate, and sonication immediately prior to NMR measurement, this ratio approaches unity. The reversibility of this behavior corroborates the presence of a pyrophosphate linker in this SCWP-peptidoglycan complex.In addition to the determination of the structure and linkage of the SCWP, a possible scenario for its biological function is discussed.