Functional aspects of the solution structure and dynamics of PAF--a highly-stable antifungal protein from Penicillium chrysogenum.
Functional aspects of the solution structure and dynamics of PAF--a highly-stable antifungal protein from Penicillium chrysogenum.
复制标题
DOI：
10.1111/j.1742-4658.2009.07011.x
复制
发表时间：
2009-05
期刊：
影响因子：
--
通讯作者：
Marx F
中科院分区：
文献类型：
--
作者：
Batta G;Barna T;Gáspári Z;Sándor S;Kövér KE;Binder U;Sarg B;Kaiserer L;Chhillar AK;Eigentler A;Leiter E;Hegedüs N;Pócsi I;Lindner H;Marx F
PAF是一种很有前途的抗真菌药物，对哺乳动物细胞没有毒性作用，因此可能成为治疗人类经常致命的曲霉感染的候选药物。PAF对敏感真菌的致病作用涉及G蛋白偶联信号转导和细胞凋亡。在这里，这种来自产黄青霉的小的阳离子抗真菌蛋白的溶液结构是通过溶液核磁共振确定的。我们证明了PAF属于其与巨型曲霉AFP最接近的同源蛋白的SCOP折叠类。PAF由五条β链组成，形成两个相互垂直排列的β片，共享一个公共界面。用核磁共振动力学和约束分子动力学计算相结合的方法，研究了三个二硫键的归属的模糊性。线索无法分辨：两个具有不同二硫键图案的乐团和一个没有S-S键的乐团展示了本质上相同的折叠。15N弛豫色散和干扰实验没有发现二硫键通过慢交换重排。测得的有序参数和3.0 ns的关联时间对于这种大小的致密单体蛋白质是合适的。我们通过定点突变证明，PAF表面高度保守且带正电荷的富含赖氨酸的结构域增强了PAF的毒性。然而，与AFP相比，PAF中OB折叠的有效性降低，这是因为较少的溶剂暴露的芳香区解释了没有壳二糖结合的原因。本工作进一步支持了对两种高度同源的抗真菌蛋白的作用模式之间已有文献记载的实质性差异的理解。
PAF is a promising antimycotic without toxic effects on mammalian cells and therefore may be a drug candidate against the often lethal Aspergillus infections in human. The pathogenesis of PAF on sensitive fungi involves G-protein coupled signaling followed by apoptosis. Here, the solution structure of this small, cationic, antifungal protein from Penicillium chrysogenum is determined by solution NMR. We proved that PAF belongs to the SCOP fold class of its closest homologue AFP from Aspergillus giganteus. PAF comprises five β—strands forming two orthogonally packed β—sheets sharing a common interface. The ambiguity in the assignment of two disulfide bonds of three was investigated by NMR dynamics combined with restrained molecular dynamics calculations. The clue could not be resolved: two ensembles with different disulfide patterns and the one with no S-S bond exhibit essentially the same fold. 15N relaxation dispersion and interference experiments did not reveal disulfide bond rearrangements via slow exchange. The measured order parameters and the 3.0 ns correlation time is appropriate for a compact monomeric protein of this size. We demonstrated by site-directed mutagenesis that the highly conserved and positively charged lysine-rich domain region on the surface enhances the toxicity of PAF. However, the efficacy of the OB fold is reduced in PAF compared to AFP, due to less solvent exposed aromatic regions explaining the absence of chitobiose binding. The present work lends further support to the understanding of the documented substantial differences between the mode of action of two highly homologous antifungal proteins.
登录
查看更多内容
影响因子：
4.8
作者：
DeSamblanx, GW;Goderis, IJ;Broekaert, WF
通讯作者：
Broekaert, WF
影响因子：
3.6
作者：
Han, KH;Seo, JA;Yu, JH
通讯作者：
Yu, JH
影响因子：
2.9
作者：
FARROW, NA;MUHANDIRAM, R;KAY, LE
通讯作者：
KAY, LE
影响因子：
2.2
作者：
GOLDMAN, M
通讯作者：
GOLDMAN, M
影响因子：
15
作者：
Kover, Katalin E.;Groves, Patrick;Batta, Gyula
通讯作者：
Batta, Gyula