Licari P J, Jarvis D L, Bailey J E
Department of Chemical Engineering, California Institute of Technology, Pasadena 91125.
Biotechnol Prog. 1993 Mar-Apr;9(2):146-52. doi: 10.1021/bp00020a005.
Four different insect cell lines that can be used as hosts for baculovirus infection were assayed for the presence of endogenous exoglycosidases. All four cell lines, derived from Spodoptera frugiperda, Trichoplusia ni, Bombyx mori, or Malacosoma disstria, contained N-acetyl-beta-glucosaminidase, N-acetyl-beta-galactosaminidase, beta-galactosidase, and sialidase activities. Exoglycosidase activities were found in cell lysates as well as cell-free supernatants from uninfected and wild-type baculovirus infected cells. Oligosaccharide analysis of cellular glycoproteins using lectins recognizing Gal beta 1, 3GalNAc, Gal beta 1, 4GlcNAc, and NeuAc alpha 2,6Gal demonstrated that only Gal beta 1,3GalNAc was present. The demonstration that these cells contain exoglycosidases raises the possibility that the oligosaccharides of baculovirus-expressed glycoproteins are subject to enzymatic degradation.
对四种可作为杆状病毒感染宿主的不同昆虫细胞系进行了内源性外切糖苷酶存在情况的检测。所有这四种细胞系分别源自草地贪夜蛾、粉纹夜蛾、家蚕或天幕毛虫,均含有N - 乙酰 - β - 氨基葡萄糖苷酶、N - 乙酰 - β - 半乳糖苷酶、β - 半乳糖苷酶和唾液酸酶活性。在未感染和野生型杆状病毒感染细胞的细胞裂解物以及无细胞上清液中均发现了外切糖苷酶活性。使用识别Galβ1,3GalNAc、Galβ1,4GlcNAc和NeuAcα2,6Gal的凝集素对细胞糖蛋白进行寡糖分析表明,仅存在Galβ1,3GalNAc。这些细胞含有外切糖苷酶这一事实增加了杆状病毒表达的糖蛋白寡糖会被酶促降解的可能性。