Cui Fengjie, Zan Xinyi, Li Yunhong, Yang Yan, Sun Wenjing, Zhou Qiang, Yu Silian, Dong Ying
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, PR China; Jiangxi Provincial Engineering and Technology Center for Food Additives Bio-production, Dexing 334221, PR China.
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, PR China.
Int J Biol Macromol. 2013 Nov;62:684-690. doi: 10.1016/j.ijbiomac.2013.10.025. Epub 2013 Oct 25.
A novel glycoprotein GFG-3a with the molecular weight of 88.01kDa and potent anti-tumor activity was isolated from the cultured mycelia of Grifola frondosa GF9801. GFG-3a was heat-sensitive with the decreasing anti-proliferative activity after treated from 56°C to 100°C for 10-120min. GFG-3a was a glycoprotein with O-glycosylation and contained 6.20% carbohydrate composed of d-arabinose, d-fructose, d-mannose, and d-glucose with a molar ratio of 1.33:4.51:2.46:1.00. FT-IR and NMR spectra proved that GFG-3a contained protein and carbohydrate portions with 3-O-methyl-galactose residues, (1→4)-linked β-galactose residues, and β-linked glucose residues. Circular dichroism (CD) revealed that GFG-3a was a predominantly β-sheet glycoprotein with a relatively small α-helical content. Protein sequencing and 3D model of GFG-3a were finally predicted by using MALDI-TOF-MS, NCBI blast search and online SWISS-MODLE Workspace service. Our findings will be a reference for the further structure-activity relationship analysis of the mushroom glycoproteins.
从灰树花GF9801的培养菌丝体中分离出一种新型糖蛋白GFG-3a，其分子量为88.01kDa，具有较强的抗肿瘤活性。GFG-3a对热敏感，在56°C至100°C处理10 - 120分钟后，其抗增殖活性降低。GFG-3a是一种O-糖基化糖蛋白，含有6.20%的碳水化合物，由d-阿拉伯糖、d-果糖、d-甘露糖和d-葡萄糖组成，摩尔比为1.33:4.51:2.46:1.00。傅里叶变换红外光谱（FT-IR）和核磁共振光谱（NMR）证明GFG-3a含有蛋白质和碳水化合物部分，其中含有3-O-甲基半乳糖残基、（1→4）连接的β-半乳糖残基和β-连接的葡萄糖残基。圆二色性（CD）显示GFG-3a是一种主要为β-折叠的糖蛋白，α-螺旋含量相对较少。最后，通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）、美国国立生物技术信息中心（NCBI）的blast搜索和在线SWISS-MODEL工作区服务预测了GFG-3a的蛋白质序列和三维模型。我们的研究结果将为进一步分析蘑菇糖蛋白的构效关系提供参考。
