Voigt J, Hinkelmann B, Harris E H
Botanisches Institut, Technische Universität Braunschweig, Germany.
Microbiol Res. 1997 Jul;152(2):189-98. doi: 10.1016/S0944-5013(97)80012-2.
Three classes of cell wall-defective mutants of the unicellular green alga Chlamydomonas reinhardtii have been described in the literature differing with respect to the amounts of cell wall material and its attachment to the plasma membrane, respectively. We have compared the production of the chaotrope-soluble cell wall polypeptides by the different mutants. These experiments have been performed by comparative Western-blot analyses using antibodies raised (1) against the deglycosylation products of the insoluble wall fraction of wild-type cells, (2) against the deglycosylation product of the '150 kDa' chaotrope-soluble cell wall polypeptide and (3) against the carbohydrate side chains of the Chlamydomonas cell wall glycoproteins, respectively. Considerably different levels of cell wall polypeptides were found in the LiCl-extracts from intact cells of the various mutant strains containing the apoplastic, chaotrope-soluble cell wall glycoproteins. No correlation was found between the amounts and the patterns of cell wall glycoproteins present in the LiCl-extracts and the electron microscopical classification of the mutant strains. All the mutant strains were shown to contain the same amounts and patterns of intracellular cell wall precursors as wild-type cells as revealed by Western-blot analyses of urea-SDS lysates of LiCl-pretreated cells. These findings indicate that the different mutant strains produce the same set of cell wall polypeptides at the same relative amounts as wild-type cells. However, in the case of some strains belonging to different classes of cell wall mutants and showing differential seggregation patterns in crosses, alterations were observed for the pattern of extracellular cell wall polypeptides present in the LiCl-extracts from intact cells and in the culture medium, respectively.
文献中描述了单细胞绿藻莱茵衣藻的三类细胞壁缺陷型突变体,它们在细胞壁物质的含量及其与质膜的附着方面存在差异。我们比较了不同突变体产生的离液剂可溶性细胞壁多肽。这些实验通过比较蛋白质免疫印迹分析进行,分别使用针对以下物质产生的抗体:(1)野生型细胞不溶性壁组分的去糖基化产物;(2)“150 kDa”离液剂可溶性细胞壁多肽的去糖基化产物;(3)莱茵衣藻细胞壁糖蛋白的碳水化合物侧链。在含有质外体、离液剂可溶性细胞壁糖蛋白的各种突变株完整细胞的LiCl提取物中,发现细胞壁多肽的水平有很大差异。在LiCl提取物中存在的细胞壁糖蛋白的量和模式与突变株的电子显微镜分类之间未发现相关性。蛋白质免疫印迹分析LiCl预处理细胞的尿素-SDS裂解物表明,所有突变株所含的细胞内细胞壁前体的量和模式与野生型细胞相同。这些发现表明,不同的突变株产生与野生型细胞相同的一组细胞壁多肽,且相对量相同。然而,对于属于不同类别的细胞壁突变体且在杂交中表现出不同分离模式的一些菌株,分别在完整细胞的LiCl提取物和培养基中存在的细胞外细胞壁多肽模式中观察到了变化。