López-Revilla R, Báez-Camargo M
Department of Cell Biology, Centro de Investigación y de Estudios Avanzados del IPN, México, DF.
Arch Med Res. 1992;23(2):95-7.
We have examined the sodium dodecyl sulfate (SDS)-induced autoproteolysis of E. invadens (PZ and IP101) and E. moshkovskii (FIC and Laredo) trophozoite lysates. Heat-treated lysates containing parahydroxy-mercuribenzoate (pHMB) of all four strains had undegraded protein patterns. Unheated pHMB-lacking lysates of PZ had two (99 and 90 kDa) and IP101 lysates had three (45, 99, 90 kDa) major proteins, whereas FIC and Laredo lysates had only one (90 kDa). Heat-treatment changed the remaining proteins to smaller ones: 37 in PZ and IP101, 33 and 37 kDa in FIC and Laredo. Unheated lysates run on gelatin-containing "substrate" gels had gelatinases whose sizes were higher in lysates containing pHMB and allowed us to detect a 200 kDa gelatinase in E. invadens strains. Our results indicate that SDS induces immediate autoproteolysis by CPs in non-histolytica trophozoites, whose proteinases appear to be processed by self-digestion, and Entamoeba proteinases vary considerably under the various conditions used to obtain and characterize them.
我们研究了十二烷基硫酸钠(SDS)诱导的侵袭内阿米巴(PZ和IP101)及莫斯科维奇内阿米巴(FIC和拉雷多)滋养体裂解物的自蛋白水解作用。所有四个菌株经热处理且含有对羟基汞苯甲酸(pHMB)的裂解物,其蛋白质模式未降解。PZ未加热且不含pHMB的裂解物有两种主要蛋白质(99 kDa和90 kDa),IP101裂解物有三种主要蛋白质(45 kDa, 99 kDa, 90 kDa),而FIC和拉雷多裂解物只有一种主要蛋白质(90 kDa)。热处理使其余蛋白质变为较小的蛋白质:PZ和IP101中为37 kDa,FIC和拉雷多中为33 kDa和37 kDa。在含明胶的“底物”凝胶上运行的未加热裂解物含有明胶酶,其大小在含pHMB的裂解物中更高,并且使我们能够在侵袭内阿米巴菌株中检测到一种200 kDa的明胶酶。我们的结果表明,SDS诱导非溶组织内阿米巴滋养体中的半胱氨酸蛋白酶立即进行自蛋白水解,其蛋白酶似乎通过自我消化进行加工,并且在用于获取和表征它们的各种条件下,内阿米巴蛋白酶有很大差异。