Okhuysen P C, DuPont H L, Sterling C R, Chappell C L
Center for Infectious Diseases, University of Texas Health Science Center-Houston Medical School 77030.
Infect Immun. 1994 Oct;62(10):4667-70. doi: 10.1128/iai.62.10.4667-4670.1994.
Cryptosporidium parvum oocysts were studied for the expression of aminopeptidase by using amino acids bound to the synthetic fluorescent substrate 7-amino-4-trifluoromethyl coumarin. After 1 h of incubation, intact oocysts showed no activity; however, homogenization and solubilization with Triton X-114 followed by phase separation yielded a 22-fold increase in aminopeptidase activity in the detergent fraction. With arginyl-6-amino-2-styrylquinoline as a substrate, aminopeptidase activity was observed in permeabilized oocysts and freshly excysted sporozoites but not on intact oocysts or empty oocyst membranes after excystation. These results suggest that C. parvum expresses an arginine aminopeptidase that is an integral protein of the sporozoite membrane.
利用与合成荧光底物7-氨基-4-三氟甲基香豆素结合的氨基酸,研究了微小隐孢子虫卵囊氨肽酶的表达。孵育1小时后,完整的卵囊未显示出活性;然而,用Triton X-114进行匀浆和增溶,随后进行相分离,去污剂组分中的氨肽酶活性增加了22倍。以精氨酰-6-氨基-2-苯乙烯基喹啉为底物,在通透的卵囊和刚脱囊的子孢子中观察到氨肽酶活性,但在完整的卵囊或脱囊后的空卵囊膜上未观察到。这些结果表明,微小隐孢子虫表达一种精氨酸氨肽酶,它是子孢子膜的一种整合蛋白。