EVIDENCE FOR ADHESIN ACTIVITY IN THE ACID-STABLE MOIETY OF THE PHOSPHOMANNOPROTEIN CELL-WALL COMPLEX OF CANDIDA-ALBICANS
EVIDENCE FOR ADHESIN ACTIVITY IN THE ACID-STABLE MOIETY OF THE PHOSPHOMANNOPROTEIN CELL-WALL COMPLEX OF CANDIDA-ALBICANS
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DOI：
10.1128/iai.62.5.1662-1668.1994
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发表时间：
1994-05-01
影响因子：
3.1
通讯作者：
CUTLER, JE
中科院分区：
文献类型：
--
作者：
KANBE, T;CUTLER, JE
先前，我们发现白色念珠菌亲水酵母细胞特异性粘附在小鼠脾边缘区巨噬细胞上。黏附素是酵母细胞壁磷酸腺苷蛋白复合物的一部分，其中一个与单克隆抗体10G发生反应的黏附素位点被鉴定为该复合物酸不稳定部分的β -1,2连接的四甘露糖。我们在此报告，在酸稳定部分的瓦复合体，以前没有报道具有粘连素活性，是在很大程度上负责酵母细胞结合到脾边缘区。采用巯基乙醇萃取和魔豆蛋白a-琼脂糖亲和层析的方法，从白念珠菌血清型B酵母细胞中分离到磷酸化蛋白复合物Fr.II。Fr.II缺乏血清a型特异性抗原因子6，该因子在酵母细胞附着于上皮细胞中起作用。通过温和的酸水解和粒度隔离分馏得到了Fr.II的酸稳定部分（即Fr.IIS）。采用魔芋豆蛋白a-琼脂糖柱层析和α -甲基- d -甘露吡喃苷线性梯度洗脱，将其进一步分离为4个组分，分别为Fr.IIS1、Fr.IIS2、Fr.IIS3和Fr.IIS4。黏附素活性的这些部分是由他们的能力，以阻止酵母细胞结合到脾边缘区。与Fr.IIS3和Fr.IIS3相比，Fr.IIS1和Fr.IIS2产生了更多的物质和更强的粘附活性。只有Fr.IIS1不与酸不稳定的-1,2-链寡糖特异性抗体（抗因子5和单克隆抗体10G）反应。fr . iis1包覆的乳胶珠以一种与酵母结合相同的模式附着在边缘区。此外，可溶性Fr.II或Fr.IIS抑制了Fr.II - 1胶乳的附着。最初的化学分析表明，Fr.IIS1上的黏附蛋白位点是一个碳水化合物，因为黏附蛋白活性被高碘酸盐氧化破坏，而不是被蛋白酶K消化破坏。
Previously, we showed that Candida albicans hydrophilic yeast cells adhere specifically to mouse splenic marginal-zone macrophages. The adhesins are part of the yeast cell wall phosphomannoprotein complex, and one adhesin site, which reacts with the monoclonal antibody 10G, was identified as a beta-1,2-linked tetramannose in the acid-labile portion of the complex. We report here that the acid-stable part of tile complex, which has not been reported previously to have adhesin activity, is in large part responsible for yeast cell binding to the splenic marginal zone. The phosphomannoprotein complex, termed Fr.II, was isolated from C. albicans serotype B yeast cells by beta-mercaptoethanol extraction and concanavalin A-agarose affinity chromatography. Fr.II is devoid of the serotype A-specific antigen factor 6, which functions in yeast cell attachment to epithelial cells. The acid-stable part of Fr.II (i.e., Fr.IIS) was obtained by mild acid hydrolysis and size exclusion fractionation. Fr.IIS was further fractionated into four fractions, Fr.IIS1, Fr.IIS2, Fr.IIS3, and Fr.IIS4, by concanavalin A-agarose column chromatography and elution with a linear gradient of alpha-methyl-D-mannopyranoside. Adhesin activity of these fractions was determined by their ability to block yeast cell binding to the splenic marginal zone. Fr.IIS1 and Fr.IIS2 yielded more material and stronger adhesin activity than either Fr.IIS3 or Fr.IIS3. Only Fr.IIS1 did not react with antibodies (anti-factor 5 and monoclonal antibody 10G) specific for the acid-labile beta-1,2-linked oligosaccharides. Fr.IIS1-coated latex beads attached specifically to the marginal zone in a pattern identical to that of yeast tell binding. Furthermore, Fr.IIS1-latex bead attachment was inhibited by soluble Fr.II or Fr.IIS. Initial chemical analyses indicate that the adhesin site on Fr.IIS1 is a carbohydrate because adhesin activity was destroyed by periodate oxidation but not by proteinase K digestion.