Delineation of interfaces on human alpha-defensins critical for human adenovirus and human papillomavirus inhibition.
Delineation of interfaces on human alpha-defensins critical for human adenovirus and human papillomavirus inhibition.
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DOI：
10.1371/journal.ppat.1004360
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发表时间：
2014-09
期刊：
影响因子：
6.7
通讯作者：
Smith JG
中科院分区：
文献类型：
--
作者：
Tenge VR;Gounder AP;Wiens ME;Lu W;Smith JG
人α-防御素是具有中和细菌和病毒靶标能力的有效抗微生物肽。单丙氨酸诱变已被用于鉴定抗菌活性和与细菌蛋白如炭疽致死因子结合的决定簇。α-防御素与无包膜病毒相互作用的类似分析有限。我们使用了一套全面的人α-防御素5（HD 5）和人中性粒细胞肽1（HNP 1）丙氨酸扫描突变体，结合和中和试验与人腺病毒（AdV）和人乳头瘤病毒（HPV）。我们已经确定了一个核心的关键的疏水残基，这是共同的决定因素，所有的病毒-防御素的相互作用进行了分析，而特异性的病毒识别是由特定的表面暴露的带电残基。疏水性残基在维持防御素的三级和四级结构以及形成用于病毒结合的界面中起多种作用。许多重要的暴露于溶剂的HD 5基团残基一起形成临界表面。然而，HNP 1没有识别出单个离散的结合面。代替整个AdV，我们在定量结合研究中使用由五邻体基底和纤维组成的重组衣壳亚基，并确定HD 5的抗病毒效力是化学计量的函数而不是亲和力。我们的研究支持了一种机制，其中α-防御素依赖于疏水和电荷-电荷相互作用以高拷贝数与这些无包膜病毒结合，以中和感染，并提供了对指导α-防御素抗病毒活性的特性的深入了解。人α-防御素是先天免疫应答的重要组成部分，并提供针对包括病毒和细菌在内的多种感染因子的初始阻断。α-防御素的抗菌活性所必需的特征已经被鉴定出来，但是我们对抗无包膜病毒活性所需的决定因素的理解是有限的。本研究利用丙氨酸扫描突变技术，系统、全面地研究了两种α-防御素的疏水性和带电残基在结合和/或中和人腺病毒和人乳头瘤病毒中的作用。我们的研究结果表明，共同的核心疏水残基是两种α-防御素抑制这些无包膜病毒的关键，每种相互作用所特有的带电残基提供了特异性。我们还发现，与病毒结合的α-防御素分子的数量与α-防御素突变体的抗病毒效力的相关性比它们对病毒衣壳的绝对亲和力更强。了解对无包膜病毒结合重要的α-防御素的共同特征将为控制这些丰富和多方面的肽在宿主防御中的功能的规则提供信息。
Human α-defensins are potent anti-microbial peptides with the ability to neutralize bacterial and viral targets. Single alanine mutagenesis has been used to identify determinants of anti-bacterial activity and binding to bacterial proteins such as anthrax lethal factor. Similar analyses of α-defensin interactions with non-enveloped viruses are limited. We used a comprehensive set of human α-defensin 5 (HD5) and human neutrophil peptide 1 (HNP1) alanine scan mutants in a combination of binding and neutralization assays with human adenovirus (AdV) and human papillomavirus (HPV). We have identified a core of critical hydrophobic residues that are common determinants for all of the virus-defensin interactions that were analyzed, while specificity in viral recognition is conferred by specific surface-exposed charged residues. The hydrophobic residues serve multiple roles in maintaining the tertiary and quaternary structure of the defensins as well as forming an interface for virus binding. Many of the important solvent-exposed residues of HD5 group together to form a critical surface. However, a single discrete binding face was not identified for HNP1. In lieu of whole AdV, we used a recombinant capsid subunit comprised of penton base and fiber in quantitative binding studies and determined that the anti-viral potency of HD5 was a function of stoichiometry rather than affinity. Our studies support a mechanism in which α-defensins depend on hydrophobic and charge-charge interactions to bind at high copy number to these non-enveloped viruses to neutralize infection and provide insight into properties that guide α-defensin anti-viral activity. Human α-defensins are an important component of the innate immune response and provide an initial block against a broad number of infectious agents, including viruses and bacteria. Characteristics of α-defensins that are necessary for their anti-bacterial activity have been identified, but our understanding of determinants required for activity against non-enveloped viruses is limited. In this work, we utilized alanine scan mutagenesis to systematically and comprehensively investigate the role of hydrophobic and charged residues of two α-defensins in binding to and/or neutralization of human adenovirus and human papillomavirus. Our results implicate common core hydrophobic residues as critical for inhibition of these non-enveloped viruses by the two α-defensins, with specificity provided by charged residues unique to each interaction. We also found that the number of α-defensin molecules bound to the virus was a stronger correlate of the anti-viral potency of the α-defensin mutants than their absolute affinity for the viral capsid. Understanding common characteristics of α-defensins important for non-enveloped virus binding will inform rules that govern the function of these abundant and multifaceted peptides in host defense.
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影响因子：
4.4
作者：
Lehrer, Robert I.;Jung, Grace;Lu, Wuyuan
通讯作者：
Lu, Wuyuan
影响因子：
4.8
作者：
Rajabi, Mohsen;de Leeuw, Erik;Lu, Wuyuan
通讯作者：
Lu, Wuyuan
DOI：
10.1111/j.1432-1033.1989.tb15008.x
发表时间：
1989-09-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
CAILLETBOUDIN, ML;STRECKER, G;MICHALSKI, JC
通讯作者：
MICHALSKI, JC
影响因子：
5.4
作者：
Snijder, Joost;Reddy, Vijay S.;Wuite, Gijs J. L.
通讯作者：
Wuite, Gijs J. L.
影响因子：
4.8
作者：
Rajabi, Mohsen;Ericksen, Bryan;Lu, Wuyuan
通讯作者：
Lu, Wuyuan