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NAOSITE : Nagasaki University's Academic Output SITE > Institute of Tropical Medicine > Bulletin > Tropical medicine > Volume 29, No. 3 >

Trypanosoma cruziの強毒株trypomastigote細胞表面には脂質様成分が優勢に存在する

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Title: Trypanosoma cruziの強毒株trypomastigote細胞表面には脂質様成分が優勢に存在する
Other Titles: Lipid-like Components Predominantly Appeared on High-virulent Trypomastigotes of Trypanosoma cruzi
Authors: 神原, 廣二 / 福間, 利英 / 中沢, 秀介 / 上村, 春樹
Authors (alternative): Kanbara, Hiroji / Fukuma, Toshihide / Nakazawa, Shusuke / Uemura, Haruki
Issue Date: 30-Sep-1987
Publisher: 長崎大学熱帯医学研究所 / Institute of Tropical Medicine, Nagasaki University
Citation: 熱帯医学 Tropical medicine 29(3). p181-185, 1987
Abstract: Trypanosoma cruziのtrypomastigote stageはこの原虫の哺乳類への感染に最も重要な役割を果す。従ってこの原虫の病原力の解明にはこのstageの原虫のもつ機能を追及するのが最も良い方法だと思われる。これまで同一株由来の病原力の異るtrypomastigoteのもつ機能の比較研究を行ってきたが、ここでは生きたtrypomastigoteがトリプシンの作用で2~3の重要な機能を失うことより、このトリプシン作用で生trypomastigoteから遊離される成分について病原力の異るものの差異を換討した。遊離成分はBenzamidine-Sepharose 6 Bのカラムを通しトリプシンを除いた後に分子量3,500以上を通過させないセルロースチューブを用いて濃縮してSDS-PAGEにて分析した。泳動後12.5% TCA固定にて強毒株由来遊離成分中に著名に多量の白濁沈殿部を認めた。この沈殿物はその後の蛋白染色では消失することから、メタノール、酢酸を含む染色溶媒または蛋白固定液中に溶解すると考えられる。更に5倍量のアセトンを用いてトリプシンによる遊離成分中の蛋白を沈殿させ、SDSを含む溶解液で溶解後SDS-PAGEにかけると、沈殿部分は消失する。これらのことから強毒株trypomastigote細胞表面には,蛋白によって結合された脂質様成分が弱毒のそれに比し著明に多量に存在することが予想される。 / The trypomastigote stage of Trypanosoma cruzi plays the most important role in completing infection of mammals. Therefore, the investigation of trypomastigote functions is useful to understand parasite virulence. We have been studying functional difference between trypomastigotes of high and low virulence. In the present work, we examined difference between components separated from living trypomastigotes of respective virulence by trypsin treatment, because trypsin treatment has been known to remove some important functions of trypomastigotes. Separated components were passed through a Benzamidine-Sepharose 6B column to remove trypsin, concentrated and dialyzed against 0.85% saline in a cellulose tube which permits molecules with less than 3,500 molecular weight (Mr) to pass, A part of concentrated sample was applied to SDS-poly-acrylamide gel electrophoresis in slab gels. The gels fixed in 12.5% trichloracetic acid after electrophoresis showed markedly larger amount of white deposits with a Mr range 37,000 to 50,000 on the lanes of high-virulent trypomastigotes than those of low-virulent. The deposits might be dissolved in methanol-acetic acid-water mixtures which were used for a staining sloution and a fixative for the following protein staining, because the part of each lane where the deposits had existed was left unstained. Furthermore, precipitated proteins of concentrated samples by five times volume of acetone showed to have lost the deposits on SDS-PAGE. These results suggested that high-virulent trypomastigotes possessed markedly larger amount of lipid-like components combined to cell surface by protein compared with low-virulent ones.
URI: http://hdl.handle.net/10069/4502
ISSN: 03855643
Type: Departmental Bulletin Paper
Appears in Collections:Volume 29, No. 3

Citable URI : http://hdl.handle.net/10069/4502

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