Repurposing Hsp104 to Antagonize Seminal Amyloid and Counter HIV Infection.
Repurposing Hsp104 to Antagonize Seminal Amyloid and Counter HIV Infection.
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DOI：
10.1016/j.chembiol.2015.07.007
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发表时间：
2015-08-20
影响因子：
--
通讯作者：
Shorter J
中科院分区：
文献类型：
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作者：
Castellano LM;Bart SM;Holmes VM;Weissman D;Shorter J
前列腺酸性磷酸酶（PAP 248 -286和PAP 85 -120）和精液凝胶（SEM 1和2）的天然蛋白水解片段在精液中形成淀粉样纤维，其捕获HIV病毒体并促进感染。例如，PAP 248 -286原纤维，称为SEVI（精液衍生的病毒感染增强剂），可以增强HIV感染几个数量级。在这里，我们设计了三种破坏性技术，通过重新利用Hsp 104（一种来自酵母的淀粉样蛋白重塑纳米机器）来快速拮抗精液淀粉样蛋白。首先，Hsp 104和增强的工程化变体Hsp 104 A503 V直接将SEVI和PAP 85 -120原纤维重塑为非淀粉样蛋白形式。其次，我们阐明了催化失活的Hsp 104支架，其不重塑淀粉样蛋白结构，但将SEVI、PAP 85 -120和SEM 1（45-107）原纤维聚集成更大的组装体。第三，我们修饰Hsp 104以与室化蛋白酶ClpP相互作用，这使得能够偶联重塑和降解以不可逆地清除SEVI和PAP 85 -120原纤维。每种策略都降低了精液淀粉样蛋白促进HIV感染的能力，并可能具有治疗效用。
Naturally occurring proteolytic fragments of prostatic acid phosphatase (PAP248-286 and PAP85-120) and semenogelins (SEM1 and 2) form amyloid fibrils in seminal fluid, which capture HIV virions and promote infection. For example, PAP248-286 fibrils, termed SEVI (Semen derived Enhancer of Viral Infection), can potentiate HIV infection by several orders of magnitude. Here, we design three disruptive technologies to rapidly antagonize seminal amyloid by repurposing Hsp104, an amyloid-remodeling nanomachine from yeast. First, Hsp104 and an enhanced engineered variant, Hsp104A503V, directly remodel SEVI and PAP85-120 fibrils into non-amyloid forms. Second, we elucidate catalytically inactive Hsp104 scaffolds that do not remodel amyloid structure, but cluster SEVI, PAP85-120, and SEM1(45-107) fibrils into larger assemblies. Third, we modify Hsp104 to interact with the chambered protease ClpP, which enables coupled remodeling and degradation to irreversibly clear SEVI and PAP85-120 fibrils. Each strategy diminished the ability of seminal amyloid to promote HIV infection and could have therapeutic utility.