Parussini F, Duschak V G, Cazzulo J J
Instituto de Investigaciones Bioquímicas Luis F. Leloir, Fundación Campomar, CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina.
Cell Mol Biol (Noisy-le-grand). 1998 May;44(3):513-9.
Cysteine proteinase isoforms, immunologically cross-reactive with cruzipain and with a similar apparent molecular mass, have been identified in epimastigotes of Trypanosoma cruzi by extraction and phase partition using the detergent Triton X-114. These isoforms are concentrated in the microsomal fraction obtained after differential centrifugation, which is known to consist essentially of plasma membrane, can be labelled by incubation of live parasites with sulfo-NHS-biotin, and bind to cystatin-sepharose affinity columns. They are present, albeit with a different electrophoretic pattern, in the epimastigote, amastigote and trypomastigote stages of the parasite.
通过使用去污剂Triton X-114进行提取和相分离,在克氏锥虫的上鞭毛体中鉴定出了与克氏锥虫蛋白酶具有免疫交叉反应且表观分子量相似的半胱氨酸蛋白酶同工型。这些同工型集中在差速离心后获得的微粒体部分,已知该部分主要由质膜组成,可通过将活寄生虫与磺基-NHS-生物素孵育进行标记,并与胱抑素-琼脂糖亲和柱结合。它们存在于寄生虫的上鞭毛体、无鞭毛体和锥鞭毛体阶段,尽管电泳图谱有所不同。