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诺氏疟原虫裂殖体感染的红细胞和正常恒河猴红细胞的唾液糖蛋白及唾液酸

Sialoglycoproteins and sialic acids of Plasmodium knowlesi schizont-infected erythrocytes and normal rhesus monkey erythrocytes.

作者信息

Howard R J, Reuter G, Barnwell J W, Schauer R

出版信息

Parasitology. 1986 Jun;92 ( Pt 3):527-43. doi: 10.1017/s0031182000065422.

Abstract

The effects of malaria infection on RBC sialic acids and sialoglycoproteins were studied with asexual blood-stage infections of Plasmodium knowlesi in rhesus monkeys. Glycoprotein radio-isotope labelling methods were used to compare the sialoglycoproteins of normal RBC and P. knowlesi schizont-infected RBC (SI-RBC). Tritiation of glycoproteins from SI-RBC with the standard sialidase + galactose oxidase/NaB3H4 method or standard periodate/NaB3H4 method was significantly decreased when compared to normal RBC. However, tritium uptake into glycoproteins was normal when SI-RBC were treated with 5-fold higher concentrations of both enzymes in the first labelling method, or with a 5-fold increase in the molar ratio of periodate to sialic acid in the second method. The mobility of tritiated host cell glycoproteins on SDS-polyacrylamide gels was identical for SI-RBC and normal RBC. New bands, possibly glycoproteins, of 230, 160, 90, 52, and 30 kDa were detected after labelling SI-RBC by the modified periodate/NaB3H4 method. Sialic acid analysis of normal rhesus monkey RBC (62 micrograms/10(10) RBC) revealed that 46% of the total sialic acid was N-glycolylneuraminic acid, 33% was N-acetyl-9-O-acetylneuraminic acid, and the remainder N-acetylneuraminic acid. SI-RBC collected either directly from infected monkeys or after in vitro culture of ring-infected RBC in horse serum, had increased total sialic acid (126 or 115 micrograms/10(10) RBC, respectively). The sialic acid content of infected RBC must increase during parasite development since RBC infected with ring-stage P. knowlesi had the same content as normal RBC. There was no significant difference in the ratio of the three sialic acids of SI-RBC and normal RBC. In contrast, the uninfected RBC from infected blood of different monkeys showed marked variation in sialic acid composition and generally had a lower sialic acid content than normal RBC.

摘要

利用恒河猴感染诺氏疟原虫无性血液期的模型,研究了疟疾感染对红细胞唾液酸和唾液糖蛋白的影响。采用糖蛋白放射性同位素标记方法,比较正常红细胞与诺氏疟原虫裂殖体感染红细胞(SI-RBC)的唾液糖蛋白。与正常红细胞相比,用标准唾液酸酶+半乳糖氧化酶/NaB3H4法或标准高碘酸盐/NaB3H4法对SI-RBC糖蛋白进行氚化标记时,标记率显著降低。然而,在第一种标记方法中,当用5倍高浓度的两种酶处理SI-RBC,或在第二种方法中使高碘酸盐与唾液酸的摩尔比增加5倍时,糖蛋白的氚摄取正常。SI-RBC和正常红细胞的氚标记宿主细胞糖蛋白在SDS-聚丙烯酰胺凝胶上的迁移率相同。用改良的高碘酸盐/NaB3H4法标记SI-RBC后,检测到230、160、90、52和30 kDa的新条带,可能为糖蛋白。对正常恒河猴红细胞(62微克/10^10个红细胞)的唾液酸分析表明,总唾液酸的46%为N-羟乙酰神经氨酸,33%为N-乙酰-9-O-乙酰神经氨酸,其余为N-乙酰神经氨酸。直接从感染猴子采集的SI-RBC或在马血清中对环状感染红细胞进行体外培养后获得的SI-RBC,总唾液酸含量均增加(分别为126或115微克/10^10个红细胞)。由于感染环状期诺氏疟原虫的红细胞与正常红细胞的唾液酸含量相同,因此感染红细胞的唾液酸含量在寄生虫发育过程中必然增加。SI-RBC和正常红细胞的三种唾液酸比例无显著差异。相比之下,来自不同猴子感染血液中的未感染红细胞在唾液酸组成上表现出明显差异,且唾液酸含量通常低于正常红细胞。

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