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白色念珠菌细胞壁含有荧光交联氨基酸二酪氨酸。

Candida albicans cell walls contain the fluorescent cross-linking amino acid dityrosine.

作者信息

Smail E H, Briza P, Panagos A, Berenfeld L

机构信息

Evans Memorial Department of Clinical Research, Boston University Medical Center Hospital, Massachusetts 02118, USA.

出版信息

Infect Immun. 1995 Oct;63(10):4078-83. doi: 10.1128/iai.63.10.4078-4083.1995.

Abstract

Several clinical and laboratory isolates of Candida albicans have a natural blue surface fluorescence when cultured and observed with sensitive optics. The localization and color of the fluorescence are similar to those of the natural fluorescence of sporulated Saccharomyces cerevisiae which is caused by the generation and surface deposition of the cross-linking amino acid dityrosine. In S. cerevisiae, dityrosine production results from the direct action of at least two genes and is responsible for resistance of the ascospores to lytic enzymes and physicochemical trauma. Among the criteria for the identification of dityrosine is pH sensitivity of the fluorescence intensity and a highly characteristic shift of the fluorescence excitation maximum with a change in pH. Video microscopy of whole Candida organisms revealed the characteristic dityrosine intensity maximum at pH approximately 10 and the intensity minimum at pH approximately 2. Separation of an acid hydrolysate of Candida cell walls by reverse-phase high-performance liquid chromatography revealed a fluorescence peak that coelutes with the reagent dityrosine. At pH approximately 10, this peak has a fluorescence excitation maximum of 320 to 325 nm, while at pH approximately 2, the excitation maximum is 285 to 290 nm. This excitation maximum shift and the observed emission maximum of approximately 410 nm are characteristic of dityrosine. Two separate strains of C. albicans were injected intraperitoneally into mice and harvested at 24 h. Blue surface fluorescence was observed, suggesting that dityrosine generation occurs in vivo as well as in vitro. This is the first report of the presence of dityrosine in a human fungal pathogen.

摘要

几种白色念珠菌的临床和实验室分离株在培养并用灵敏光学仪器观察时具有天然的蓝色表面荧光。荧光的定位和颜色与产孢酿酒酵母的天然荧光相似,后者是由交联氨基酸二酪氨酸的生成和表面沉积引起的。在酿酒酵母中,二酪氨酸的产生是至少两个基因直接作用的结果,并且负责子囊孢子对裂解酶和物理化学损伤的抗性。鉴定二酪氨酸的标准包括荧光强度的pH敏感性以及随着pH变化荧光激发最大值的高度特征性变化。对完整念珠菌生物体的视频显微镜观察显示,在pH约为10时二酪氨酸强度最大,在pH约为2时强度最小。通过反相高效液相色谱法分离念珠菌细胞壁的酸水解产物,发现一个与试剂二酪氨酸共洗脱的荧光峰。在pH约为10时,该峰的荧光激发最大值为320至325nm,而在pH约为2时,激发最大值为285至290nm。这种激发最大值的变化以及观察到的约410nm的发射最大值是二酪氨酸的特征。将两株不同的白色念珠菌菌株腹腔注射到小鼠体内,并在24小时后收获。观察到蓝色表面荧光,表明二酪氨酸的生成在体内和体外均会发生。这是关于人类真菌病原体中存在二酪氨酸的首次报道。

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