Proteomic analysis of proteins released from growth-arrested Candida albicans following exposure to caspofungin
Proteomic analysis of proteins released from growth-arrested Candida albicans following exposure to caspofungin
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DOI：
10.3109/13693780903405782
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发表时间：
2010-06-01
期刊：
影响因子：
2.9
通讯作者：
Kavanagh, Kevin
中科院分区：
文献类型：
--
作者：
Kelly, Judy;Kavanagh, Kevin
棘白菌素（E.例如，在一个实施例中，卡泊芬净）是一类相对较新的抗真菌药物，其通过抑制细胞壁中β-1，3-葡聚糖的合成而起作用，从而导致细胞溶解。在这项工作中，监测卡泊芬净对暴露于药物的白色念珠菌酵母的非生长细胞释放肽的影响。暴露于0.19 μ g/ml卡泊芬净导致从细胞中释放氨基酸和小分子量和大分子量蛋白质，如1-和2-维凝胶电泳所示。基质辅助激光解吸/电离飞行时间质谱（MALDI-ToF）质谱法被用来鉴定一些逃逸的肽，这些肽在暴露于药物后强度增加。一些壁相关蛋白（e。例如，在一个实施例中，磷酸甘油酸激酶）和许多糖酵解酶（磷酸甘油酸激酶1，果糖-二磷酸醛缩酶）。重要的是，一些释放的蛋白质（e。例如，在一个实施例中，丙酮酸激酶、烯醇化酶1、磷酸甘油酸脱氢酶、甘油醛3-磷酸脱氢酶、果糖二磷酸醛缩酶和醇脱氢酶1）在本质上是高度免疫原性的。结果表明，不生长的C。白色念珠菌细胞对卡泊芬净的作用敏感，并且卡泊芬净介导的蛋白质从这些细胞中的释放可导致更强的体内免疫应答。这份报告说明，除了阻碍细胞壁的合成，卡泊芬净也可能干扰真菌细胞壁的渗透性。
The echinocandins (e. g., caspofungin) are a relatively new class of antifungal drugs that function by inhibiting the synthesis of beta-1,3-glucan in the cell wall and thus lead to lysis of the cell. In this work the effect of caspofungin on the release of peptides from non-growing cells of the yeast Candida albicans that had been exposed to the drug was monitored. Exposure to 0.19 mu g/ml caspofungin resulted in the release of amino acids from cells and of both small and large molecular weight proteins as demonstrated by 1- and 2-dimensional gel electrophoresis. Matrix-assisted laser desorption/ionizationtime of flight-mass spectrometry (MALDI-ToF) mass spectrometry was employed to identify a number of escaped peptides that were found to have increased in intensity upon exposure to the drug. A number of wall-associated proteins (e. g., phosphoglycerate kinase) and a number of glycolytic enzymes (phosphoglycerate mutase 1, fructose-bisphosphate aldolase) were identified. Importantly, several released proteins (e. g., pyruvate kinase, enolase 1, phosphoglycerate mutase, glyceraldehydes 3-phosphate dehydrogenase, fructose bisphosphate aldolase and alcohol dehydrogenase 1) are highly immunogenic in nature. The results presented here demonstrate that non-growing C. albicans cells are susceptible to the effect of caspofungin and that the caspofungin-mediated release of proteins from such cells could lead to a stronger immune response in vivo. This report illustrates that, in addition to hampering cell wall synthesis, caspofungin may also interfere with the permeability of the fungal cell wall.