TRANSGLUTAMINASE AND SALIVA IN ORAL CANDIDA ADHESION

转谷氨酰胺酶和唾液在口腔念珠菌粘附中的作用

• 批准号: 2131135
• 负责人: STEVEN D BRADWAY
• 金额: $ 10.09万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2131135
• 关键词: Candida albicans; Saccharomyces cerevisiae; active sites; amylases; artificial chromosomes; blocking antibody; candidiasis; catalyst; cell adhesion; cell adhesion molecules; cell membrane; cell wall; complementary DNA; covalent bond; crosslink; enzyme inhibitors; enzyme linked immunosorbent assay; enzyme mechanism; epithelium; genetic library; genetic strain; human subject; laboratory mouse; laboratory rabbit; monoclonal antibody; protein glutamine gamma glutamyltransferase; protein purification; protein structure function; saliva; site directed mutagenesis; tissue /cell culture; transfection /expression vector

C. albicans infection i.e. candidiasis, produces recurrent, invasive oral lesions in a steadily increasing population of immunocompromised patients. The necessity of using toxic, systemic antimycotics in these patients has produced a renewed interest in developing alternative therapies for the treatment of candidiasis. Adhesion to epithelial cells, the first event in oral candidiasis, is mediated by a repertoire of adhesive molecules expressed on the surface of C. albicans cell wall. Previous studies suggests that the temporal escalation of non-covalent bond formation between candida adhesive molecules and epithelial cell surface molecules initially elicits a reversible adhesion which is quickly stabilized to become functionally irreversible. Additionally, saliva has been proposed to modulated candida adherence. These studies, however, have focused on characterization of C. albicans adhesive molecules; the host contribution to adhesion, i.e. the role of saliva and epithelial cell factors, have only been superficially characterized. Preliminary experiments in our laboratory indicate that an epithelial surface enzyme, transglutaminase, covalently cross-links inducible, morphotype specific, C. albicans cell wall protein to proteins on epithelial cell surfaces. [Additionally, the salivary proteins (proline-rich proteins > cystatin > amylase) react with transglutaminase and therefore, may compete with candida proteins for reactivity with transglutaminase.] Thus, we hypothesize that salivary proteins may modulate a transglutaminase catalyzed, covalent mechanism of adhesion by competing with candida cell wall proteins for transglutaminase catalyzed cross-linking to the epithelial cell surface. The objectives of this application are to confirm this hypothesis by utilizing the following specific aims: 1) Characterize the role of epithelial transglutaminase in stabilizing C. albicans adhesion to buccal epithelial cells; [2) Characterize the effect of salivary proteins on transglutaminase catalyzed mechanisms of adhesion; and 3) Characterize C. albicans transglutaminase reactive proteins (TRP) by transformation of non-adherent S. cerevisiae with TRP-cDNA: ] To our knowledge this is the first study of a covalent mechanisms in microbial adhesion. Conformation and characterization of this mechanism will provide knowledge adjunctive to the study of C. albicans adhesion. The results of this study may lead to alternative therapies for the treatment of candidal infection in immunocompromised patients.

白色念珠菌感染，即念珠菌病，会产生复发性、侵袭性口腔疾病 免疫功能低下人群中的病变不断增加 患者。 在这些情况下使用有毒的、系统性抗真菌药的必要性 患者对开发替代品产生了新的兴趣 治疗念珠菌病的疗法。 与上皮的粘附 细胞，口腔念珠菌病的第一个事件，是由一套系统介导的 白色念珠菌细胞壁表面表达的粘附分子。 先前的研究表明，非共价键的时间升级 念珠菌粘附分子与上皮细胞之间的键形成 表面分子最初引起可逆粘附，即 很快稳定下来，变得功能上不可逆。 此外， 有人提出唾液可以调节念珠菌的粘附。 这些研究， 然而，重点关注白色念珠菌粘合剂的表征 分子；宿主对粘附的贡献，即唾液和 上皮细胞因子，仅得到了表面特征。 我们实验室的初步实验表明，上皮细胞 表面酶、转谷氨酰胺酶、共价交联诱导型、 形态型特异性，白色念珠菌细胞壁蛋白至蛋白质 上皮细胞表面。 [此外，唾液蛋白 （富含脯氨酸的蛋白质 > 半胱氨酸蛋白酶抑制剂 > 淀粉酶）与转谷氨酰胺酶发生反应 因此，可能与念珠菌蛋白竞争反应性 ]因此，我们假设唾液蛋白可能 调节转谷氨酰胺酶催化的共价粘附机制 与念珠菌细胞壁蛋白竞争转谷氨酰胺酶催化 交联至上皮细胞表面。 本次活动的目标 应用程序将通过利用以下内容来证实这一假设 具体目标： 1) 表征上皮转谷氨酰胺酶在稳定 C 中的作用。 白色念珠菌粘附于颊上皮细胞； [2) 表征效果 唾液蛋白对转谷氨酰胺酶催化机制的影响 附着力； 3) 表征白色念珠菌转谷氨酰胺酶反应性 蛋白质（TRP）通过转化非贴壁酿酒酵母 TRP-cDNA：] 据我们所知，这是首次对共价机制进行研究 微生物粘附。 该机制的构象和表征 将为白色念珠菌粘附的研究提供辅助知识。 这项研究的结果可能会带来替代疗法 治疗免疫功能低下患者的念珠菌感染。