CHARACTERIZATION OF ESTROGEN BINDING PROTEIN FROM CANDIDA ALBICANS

白色念珠菌雌激素结合蛋白的表征

• 批准号: 6308857
• 负责人: SUSAN M MILLER
• 金额: $ 0.99万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6308857
• 关键词: AIDS; biological products; biomedical resource; clinical research; endocrine gland /system; enzymes; hormones; immunology; infection; inflammation; lymphatic system; microorganism; plants; spectrometry; structural biology; virus; women's health

Candida albicans is the most commonly occurring opportunistic fungal pathogen in humans. It has recently been shown that when estrogens are applied to cultures of C. albicans, the cells undergo a morphological change to the more infectious form of the organism. Based on thislead, an estrogen-binding protein from C. albicans has been isolated and cloned by a group at Stanford. The protein shows 46% sequence identity to a flavin-containing redox protein from Saccharomyces cerevisiae that has been studied for some years, but for which the function is still not known. We have begun a collaboration to characterize the scope of catalytic activity, redox properties and ligand binding behavior of this protein from C. albicans, with the eventual goal of defining its true function and possible relationship to infectivity. We are currently receiving protein from our collaborators at Stanford that has been expressed as a fusion protein in E. coli and proteolytically clipped after purification. In order to compare the properties of this recombinant protein with wild type protein, we have purified enzyme from a constitutively expressing strain of Candida and have shown it to have similar activity and molecular weight properties. Before proceeding with detailed characterization, we would like to determine whether the "wild type" protein we have isolated is indeed the same as the recombinant. In addition, there are at least two similar genes for the homologous protein in Saccharomyces and our collaborators have indicated that they have also cloned a second gene in Candida. Thus, we may have a misture of isozymes in the purified sample. At this point we would like to determine whether we have isolated a single protein or a mixture of isozymes. Then in addition, we want to obtain and compare sequence data for both recombinant and "wild type" proteins to verify that we have isolated the correct protein.

白色念珠菌是最常见的机会性 人体内的真菌病原体 最近的研究表明，当 雌激素应用于C.白色念珠菌，细胞经历 形态学上的变化，以更具感染性的形式的有机体。 基于这一线索，从C.白色念珠菌具有 被斯坦福大学的一个研究小组分离并克隆。 蛋白质显示 46%的序列同一性，含黄素的氧化还原蛋白，从 酿酒酵母，已经研究了一些年，但对于 其功能仍然未知。 我们开始了一项合作 表征催化活性、氧化还原性质和 该蛋白质的配体结合行为。白色念珠菌， 确定其真正功能和可能关系的最终目标 传染性。 我们目前正在从我们的 斯坦福大学的合作者，已经表达为融合蛋白 在大肠大肠杆菌中，并在纯化后进行蛋白水解剪切。 为了 为了比较该重组蛋白与野生型的性质， 蛋白质，我们已经从组成型表达的 并已显示其具有类似活性， 分子量性质 在进行详细的 表征，我们想确定是否“野生型” 我们分离的蛋白质确实与重组体相同。 在 此外，至少有两个相似的基因的同源 我们的合作者已经表明， 他们还在念珠菌中克隆了第二个基因。 因此，我们可能有一个 纯化样品中同工酶的减少。 在这一点上，我们将 我想确定我们是否已经分离出一种单一的蛋白质或 同工酶的混合物。 此外，我们还想获得并比较 重组蛋白和“野生型”蛋白的序列数据， 我们已经分离出了正确的蛋白质