N-Methylation as a Strategy for Enhancing the Affinity and Selectivity of RNA-binding Peptides: Application to the HIV-1 Frameshift-Stimulating RNA.
N-Methylation as a Strategy for Enhancing the Affinity and Selectivity of RNA-binding Peptides: Application to the HIV-1 Frameshift-Stimulating RNA.
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DOI：
10.1021/acschembio.5b00682
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发表时间：
2016-01-15
影响因子：
4
通讯作者：
Miller BL
中科院分区：
文献类型：
--
作者：
Hilimire TA;Bennett RP;Stewart RA;Garcia-Miranda P;Blume A;Becker J;Sherer N;Helms ED;Butcher SE;Smith HC;Miller BL
人类免疫缺陷病毒（HIV）1型使用−1程序性核糖体移码（−1 PRF）事件来翻译其酶，该酶来自用于编码病毒结构蛋白的相同转录本。该事件的频率受到高度管制，并且已证明与正常5-10%频率的显著偏离可降低病毒感染性。移码主要由移码刺激信号RNA（FSS-RNA）调节，FSS-RNA是一种生物学稳定的高度保守的茎环，已被提议作为治疗靶点。我们描述了一系列能够以低纳摩尔亲和力和高选择性结合HIV-1 FSS RNA茎环的N-甲基肽的设计、合成和测试。表面等离子体共振（SPR）数据表明增加的亲和力是基本上增强的接通速率的反映。在假型病毒试验中，化合物容易穿透细胞膜并抑制HIV感染性。病毒感染性抑制与病毒颗粒中Gag和Gag-Pol比例的化合物依赖性变化相关。作为第一个化合物，具有单位数纳摩尔亲和力的FSS RNA和抑制细胞中的HIV的能力，这些研究支持使用N-甲基化来增强RNA结合肽的亲和力，选择性和生物活性。
Human Immunodeficiency Virus (HIV) type 1 uses a −1 programmed ribosomal frameshift (−1 PRF) event to translate its enzymes from the same transcript used to encode the virus’ structural proteins. The frequency of this event is highly regulated, and significant deviation from the normal 5–10% frequency has been demonstrated to decrease viral infectivity. Frameshifting is primarily regulated by the Frameshift Stimulatory Signal RNA (FSS-RNA), a thermodynamically stable, highly conserved stem loop that has been proposed as a therapeutic target. We describe the design, synthesis, and testing of a series of N-methyl peptides able to bind the HIV-1 FSS RNA stem loop with low nanomolar affinity and high selectivity. Surface plasmon resonance (SPR) data indicates increased affinity is a reflection of a substantially enhanced on rate. Compounds readily penetrate cell membranes and inhibit HIV infectivity in a pseudotyped virus assay. Viral infectivity inhibition correlates with compound-dependent changes in the ratios of Gag and Gag-Pol in virus particles. As the first compounds with both single digit nanomolar affinities for the FSS RNA and an ability to inhibit HIV in cells, these studies support the use of N-methylation for enhancing the affinity, selectivity, and bioactivity of RNA-binding peptides.