Expression and Characterization of Cryptococcus neoformans Recombinant App1

被引：4
|
作者：
Qureshi, Asfia [1 ]
Williams, Virginia [2 ]
Del Poeta, Maurizio [1 ,2 ,3 ,4 ]
机构：
[1] Med Univ S Carolina, Dept Biochem & Mol Biol, Charleston, SC 29425 USA
[2] Med Univ S Carolina, Dept Microbiol & Immunol, Charleston, SC 29425 USA
[3] Med Univ S Carolina, Dept Craniofacial Biol, Charleston, SC 29425 USA
[4] Med Univ S Carolina, Div Infect Dis, Charleston, SC 29425 USA
来源：
MYCOPATHOLOGIA | 2012年 / 173卷 / 5-6期
基金：
美国国家卫生研究院;
关键词：
Cryptococcus neoformans; Antiphagocytic protein 1 (App1); Intermolecular disulfide bridges; Insect cells; Lentivirus; PROTEIN SECONDARY STRUCTURE; MELANIN; VIRULENCE; DICHROISM;
D O I：
10.1007/s11046-011-9486-7
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
We characterized Cryptococcus neoformans recombinant antiphagocytic protein 1 (rApp1) by SDS-PAGE, gel filtration chromatography, circular dichroism, and fluorescence spectroscopy. rApp1 produced by C. neoformans var. grubii contains an odd number of cysteines; therefore, it has the ability to form intermolecular disulfide bridges which can lead to the formation of amyloid fibrils in the absence of beta-mercaptoethanol or DTT in vitro. Alternate approaches to over-expression of rApp1 in the Lepidopteran High Five((TM)) Insect cell line using pIZ/V5-His and in lentivirus were explored and are described. Finally, localization of App1 in vivo was examined in the presence and absence of the capsule.
引用
收藏
页码：395 / 405
页数：11
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