Protein Research Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City of Scientific Research and Technology Applications (SRTA-City), New Borg EL Arab 21934, Alexandria, Egypt; Botany & Microbiology Department, Faculty of Science, Al Azhar University (Girls Branch), Cairo, Egypt.
Botany & Microbiology Department, Faculty of Science, Al Azhar University (Girls Branch), Cairo, Egypt.
Int J Biol Macromol. 2022 Jun 15;210:415-429. doi: 10.1016/j.ijbiomac.2022.04.120. Epub 2022 Apr 30.
In our survey for a new antiviral agent, two types of lectin were purified from Nostoc muscorum using both ion-exchange and affinity columns chromatography. Nostoc muscorum lectins (NMLs) are categorized based on their carbohydrate preference. Nostoc muscorum lectin-1(NML-1) exhibited a strict binding specificity for complex glycoproteins without linked carbohydrates, and the other displayed specificity for α- glycosides mannose polymers (NML-2) and was classified as a glycoprotein with 16.8% linked carbohydrates. NML-1 displayed a single band of 166 kDa on native-PAGE and two bands of 81 kDa and 85 kDa on SDS-PAGE, which confirmed the heterodimeric nature of this lectin. While NML-2 is a 50 kDa glycoprotein composed of 25 kDa subunits. Physical characterization of NML-1 displayed its stability at a higher temperature of 90 °C for 5 min and over a wide pH range (4-9), while MNL-2 displayed stability up to a temperature of 80 °C for 25 min and a pH range of 5-8. NML-1 didn't require metal ions for agglutination activity, while the activity of NML-2 was doubled by manganese ions. The antiviral activity of two lectins was assessed against herpes simplex type-1 (HSV-1) using a plaque assay which revealed that NML-1 inhibited HSV-1 infection at an early stage in contrast to NML-2 which exerted its antiviral effect at the late stage of infection. These results suggest that Nostoc muscorum is a unique lead for antiviral drug discovery as it is a novel source for antiviral lectins with different modes of action.
在我们寻找新型抗病毒药物的研究中,使用离子交换和亲和层析柱从地木耳中纯化了两种凝集素。地木耳凝集素(NMLs)根据其碳水化合物偏好进行分类。地木耳凝集素-1(NML-1)对无连接碳水化合物的复杂糖蛋白表现出严格的结合特异性,而另一种则对α-糖苷甘露聚糖聚合物(NML-2)表现出特异性,被归类为具有 16.8%连接碳水化合物的糖蛋白。NML-1 在天然-PAGE 上显示出 166 kDa 的单一条带,在 SDS-PAGE 上显示出 81 kDa 和 85 kDa 的两条带,这证实了该凝集素的异二聚体性质。而 NML-2 是一种 50 kDa 的糖蛋白,由 25 kDa 的亚基组成。NML-1 的物理特性显示其在 90°C 下稳定 5 分钟,且在较宽的 pH 范围(4-9)下稳定,而 MNL-2 在 80°C 下稳定 25 分钟,pH 范围为 5-8。NML-1 不需要金属离子即可发挥凝集活性,而 NML-2 的活性在锰离子存在下增加了一倍。两种凝集素的抗病毒活性通过噬菌斑测定法评估了对单纯疱疹病毒-1(HSV-1)的抑制作用,结果表明 NML-1 抑制 HSV-1 感染发生在早期,而 NML-2 则在感染晚期发挥其抗病毒作用。这些结果表明,地木耳是抗病毒药物发现的独特先导化合物,因为它是具有不同作用模式的新型抗病毒凝集素的来源。