Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical Sensing.
Structurally Different Yet Functionally Similar: Aptamers Specific for the Ebola Virus Soluble Glycoprotein and GP1,2 and Their Application in Electrochemical Sensing.
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DOI：
10.3390/ijms24054627
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发表时间：
2023-02-27
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
--
作者：
埃博拉病毒糖蛋白（GP）基因模板几个mRNA，产生病毒体相关的跨膜蛋白或两个分泌的糖蛋白之一。可溶性糖蛋白（sGP）是主要产物。GP 1和sGP共享295个氨基酸的氨基末端序列，但四级结构不同，GP 1是异源六聚体，GP 2和sGP是同源二聚体。针对sGP选择两种结构不同的DNA适体，其也结合GP 1，2。将这些DNA适体与2′FY-RNA适体进行比较，以确定它们与埃博拉GP基因产物的相互作用。这三种适体在溶液中和病毒体上对sGP和GP 1，2具有几乎相同的结合等温线。它们表现出对sGP和GP 1，2的高亲和力和选择性。此外，一个适体，作为传感元件在电化学格式，检测GP 1，2假型病毒体和sGP在血清的存在下，包括从埃博拉病毒感染的猴子具有高灵敏度。我们的研究结果表明，适体与sGP相互作用的单体之间的界面，这是从大多数抗体结合的蛋白质上的网站不同。三种结构不同的适体在功能特征上的显著相似性表明，适体与抗体一样，在蛋白质上具有优选的结合位点。
The Ebola virus glycoprotein (GP) gene templates several mRNAs that produce either the virion-associated transmembrane protein or one of two secreted glycoproteins. Soluble glycoprotein (sGP) is the predominant product. GP1 and sGP share an amino terminal sequence of 295 amino acids but differ in quaternary structure, with GP1 being a heterohexamer with GP2 and sGP a homodimer. Two structurally different DNA aptamers were selected against sGP that also bound GP1,2. These DNA aptamers were compared with a 2′FY-RNA aptamer for their interactions with the Ebola GP gene products. The three aptamers have almost identical binding isotherms for sGP and GP1,2 in solution and on the virion. They demonstrated high affinity and selectivity for sGP and GP1,2. Furthermore, one aptamer, used as a sensing element in an electrochemical format, detected GP1,2 on pseudotyped virions and sGP with high sensitivity in the presence of serum, including from an Ebola-virus-infected monkey. Our results suggest that the aptamers interact with sGP across the interface between the monomers, which is different from the sites on the protein bound by most antibodies. The remarkable similarity in functional features of three structurally distinct aptamers suggests that aptamers, like antibodies, have preferred binding sites on proteins.
影响因子：
4.5
作者：
Alfson KJ;Goez-Gazi Y;Gazi M;Staples H;Mattix M;Ticer A;Klaffke B;Stanfield K;Escareno P;Keiser P;Griffiths A;Chou YL;Niemuth N;Meister GT;Cirimotich CM;Carrion R Jr
通讯作者：
Carrion R Jr
影响因子：
3.7
作者：
Shedlock DJ;Bailey MA;Popernack PM;Cunningham JM;Burton DR;Sullivan NJ
通讯作者：
Sullivan NJ
影响因子：
4.6
作者：
Shubham S;Hoinka J;Banerjee S;Swanson E;Dillard JA;Lennemann NJ;Przytycka TM;Maury W;Nilsen-Hamilton M
通讯作者：
Nilsen-Hamilton M