Matsubara Teruhiko, Shibata Rabi, Sato Toshinori
Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University Yokohama, Japan.
Front Microbiol. 2016 Apr 7;7:468. doi: 10.3389/fmicb.2016.00468. eCollection 2016.
Hemagglutinin (HA) plays an important role in the first step of influenza virus (IFV) infection because it initiates the binding of the virus to the sialylgalactose linkages of the receptors on the host cells. We herein demonstrate that a HA-binding peptide immobilized on a solid support is available to bind to HA and IFV. We previously obtained a HA-binding pentapeptide (Ala-Arg-Leu-Pro-Arg), which was identified by phage-display selection against HAs from random peptide libraries. This peptide binds to the receptor-binding site of HA by mimicking sialic acid. A peptide-conjugated lipid (pep-PE) was chemically synthesized from the peptide and a saturated phospholipid. A lipid bilayer composed of pep-PE and an unsaturated phospholipid (DOPC) was immobilized on a mica plate; and the interaction between HA and the pep-PE/DOPC membrane was investigated using atomic force microscopy. The binding of IFV to the pep-PE/DOPC membrane was detected by an enzyme-linked immunosorbent assay and real-time reverse transcription PCR. Our results indicate that peptide-conjugated lipids are a useful molecular device for the detection of HA and IFV.
血凝素（HA）在流感病毒（IFV）感染的第一步中起着重要作用，因为它启动了病毒与宿主细胞表面受体的唾液酸半乳糖连接的结合。我们在此证明，固定在固体支持物上的HA结合肽可用于结合HA和IFV。我们之前获得了一种HA结合五肽（Ala-Arg-Leu-Pro-Arg），它是通过针对来自随机肽库的HA进行噬菌体展示筛选而鉴定出来的。该肽通过模拟唾液酸与HA的受体结合位点结合。由该肽和饱和磷脂化学合成了一种肽缀合脂质（pep-PE）。将由pep-PE和不饱和磷脂（DOPC）组成的脂质双层固定在云母片上；并使用原子力显微镜研究HA与pep-PE/DOPC膜之间的相互作用。通过酶联免疫吸附测定和实时逆转录PCR检测IFV与pep-PE/DOPC膜的结合。我们的结果表明，肽缀合脂质是检测HA和IFV的有用分子装置。
