Schauer R, Wember M, Howard R J
Hoppe Seylers Z Physiol Chem. 1984 Feb;365(2):185-94. doi: 10.1515/bchm2.1984.365.1.185.
The capacity of Plasmodia to synthesize sialic acids was investigated by adding radioactive acetate to short-term in vitro cultures of the intraerythrocytic asexual forms of three malaria parasites (the human malaria Plasmodium falciparum in Aotus trivirgatus erythrocytes; the simian malaria P. knowlesi in rhesus monkey erythrocytes; the rodent malaria P. berghei in mouse erythrocytes) and to cultures of extracellular zygotes of the avian malaria P. gallinaceum. Radioactive acetate was added to normal rhesus monkey erythrocytes and to cells of the murine myeloma NS-1 for comparison. Although [1-14C]-acetate labeled many proteins with each malaria parasite and the NS-1 cells, analysis of purified sialic acids revealed that only with the NS-1 cells was radioactivity incorporated into sialic acids. Furthermore, N-acetyl[6-3H]mannosamine was not incorporated into sialic acids or malarial glycoproteins when added to P. knowlesi cultures. All of the malaria parasites underwent growth or differentiation during these experiments as measured by [35S]methionine uptake into protein and by light microscopy. Extracellular parasites largely free of erythrocyte membranes were prepared to determine whether Plasmodia contain sialic acids that are not labeled by exogenous precursors. Purified merozoites of P. knowlesi and zygotes of P. gallinaceum did not contain detectable amounts of sialic acids on chemical analysis. Thus, although we could show that Plasmodia can incorporate radioactive sugars such as glucosamine, galactose and mannose into proteins, presumably glycoproteins, they do not synthesize sialic acids or sialo-glycoproteins, nor do they contain sialo-glycoconjugates of host origin.
通过向三种疟原虫红细胞内无性体的短期体外培养物(三带犰狳红细胞内的人类疟原虫恶性疟原虫;恒河猴红细胞内的猿猴疟原虫诺氏疟原虫;小鼠红细胞内的啮齿动物疟原虫伯氏疟原虫)以及鸡疟原虫细胞外合子培养物中添加放射性乙酸盐,研究疟原虫合成唾液酸的能力。将放射性乙酸盐添加到正常恒河猴红细胞和鼠骨髓瘤NS-1细胞中作为对照。尽管[1-¹⁴C]-乙酸盐标记了每种疟原虫和NS-1细胞的许多蛋白质,但对纯化唾液酸的分析表明,只有NS-1细胞的放射性被掺入到唾液酸中。此外,当添加到诺氏疟原虫培养物中时,N-乙酰[6-³H]甘露糖胺未掺入唾液酸或疟原虫糖蛋白中。在这些实验中,通过[³⁵S]甲硫氨酸掺入蛋白质以及光学显微镜观察,所有疟原虫都经历了生长或分化。制备了基本不含红细胞膜的细胞外寄生虫,以确定疟原虫是否含有未被外源前体标记的唾液酸。化学分析显示,纯化的诺氏疟原虫裂殖子和鸡疟原虫合子不含可检测量的唾液酸。因此,尽管我们可以证明疟原虫能够将放射性糖如葡糖胺、半乳糖和甘露糖掺入蛋白质(可能是糖蛋白)中,但它们不合成唾液酸或唾液酸糖蛋白,也不含有宿主来源的唾液酸糖缀合物。