Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Street 16/10, 117997 Moscow, Russia.
Protein Eng Des Sel. 2013 Sep;26(9):547-52. doi: 10.1093/protein/gzt034. Epub 2013 Jul 19.
Hemagglutinin (HA), the trimeric spike of influenza virus, catalyzes fusion of viral and cellular membranes. We have synthesized the anchoring peptide including the linker, transmembrane region and cytoplasmic tail (HA-TMR-CT) in a cell-free system. Furthermore, to mimic the palmitoylation of three conserved cysteines within the CT, we chemically alkylated HA-TMR-CT using hexadecyl-methanethiosulfonate. While the nuclear magnetic resonance spectroscopy showed pure and refolded peptides, the formation of multiple oligomers of higher order impeded further structural analysis. Circular dichroism spectroscopy of both alkylated and non-alkylated HA-TMR-CT revealed an α-helical secondary structure. No major impact of the fatty acids on the secondary structure was detected.
血凝素 (HA) 是流感病毒的三聚体刺突，能催化病毒和细胞膜融合。我们在无细胞体系中合成了锚定肽，包含接头、跨膜区和细胞质尾巴（HA-TMR-CT）。此外，为了模拟 CT 中三个保守半胱氨酸的棕榈酰化，我们使用十六烷基甲硫磺酸对 HA-TMR-CT 进行了化学烷基化。虽然核磁共振光谱显示了纯的和重折叠的肽，但高序的多种寡聚物的形成阻碍了进一步的结构分析。烷基化和非烷基化的 HA-TMR-CT 的圆二色性光谱显示出 α-螺旋二级结构。未检测到脂肪酸对二级结构有重大影响。
