Ramon A M, Montero M, Sentandreu R, Valentin E
Sección De Microbiología, Facultad De Farmacia, Universidad De Valencia, Spain.
Res Microbiol. 1999 Mar;150(2):95-103. doi: 10.1016/s0923-2508(99)80027-8.
Linkages of Ywp1 to other components of the Yarrowia lipolytica mycelial cell wall were studied by extraction with beta-mercaptoethanol and zymolyase (a beta-glucanase complex) and by the use of rabbit polyclonal antibody preparation raised against Ywp1. Ywp1 complexed with an N-glycosylated cell wall protein(s) to form supramolecular complexes through disulphide bridges (extractable with beta-mercaptoethanol) or bonded to beta-1,3-glucan (extractable with zymolyase). The lack of a specific morphological phenotype when YWP1 was knocked out by gene disruption might indicate that other proteins present in the cell wall of Y. lipolytica compensated for its loss. In this mutant, the electrophoretic pattern of proteins, detected with polyclonal antibodies against the entire cell wall, was different from that obtained with the parental strain, but sensitivity to calcofluor white, zymolyase and chitinase did not change. Quantitative analysis of fluorescence emitted by cells in the presence of fluorescent wheat germ agglutinin (FITC-WGA) indicated that chitin was organized in the cell wall of the mutant cells in a form different from that in the parental strain.
通过用β-巯基乙醇和溶壁酶（一种β-葡聚糖酶复合物）提取以及使用针对Ywp1制备的兔多克隆抗体制剂，研究了解脂耶氏酵母菌丝细胞壁中Ywp1与其他成分的联系。Ywp1与一种N-糖基化细胞壁蛋白形成超分子复合物，通过二硫键（可用β-巯基乙醇提取）连接，或与β-1,3-葡聚糖结合（可用溶壁酶提取）。当通过基因破坏敲除YWP1时缺乏特定的形态表型，这可能表明解脂耶氏酵母细胞壁中存在的其他蛋白质补偿了它的缺失。在这个突变体中，用针对整个细胞壁的多克隆抗体检测到的蛋白质电泳图谱与亲本菌株不同，但对荧光增白剂、溶壁酶和几丁质酶的敏感性没有变化。在荧光小麦胚凝集素（FITC-WGA）存在下对细胞发出的荧光进行定量分析表明，几丁质在突变体细胞壁中的组织形式与亲本菌株不同。
