Inocencio N M, Moehring J M, Moehring T J
Department of Microbiology and Molecular Genetics, Markey Center for Molecular Genetics, University of Vermont, Burlington 05405.
J Biol Chem. 1994 Dec 16;269(50):31831-5.
We have demonstrated that the native proenzymatic form of Pseudomonas exotoxin A can be cleaved at its specific activation site by furin in intact Chinese hamster ovary cells or in vitro by furin in isolated membrane fractions from these cells. We have compared the activity of furin in cell membrane fractions with that of purified, recombinant human furin. We have verified that RPE.40, a Pseudomonas toxin-resistant mutant cell strain, is mutant in the fur gene, and we have demonstrated that these cells are deficient in cleavage of the toxin. We have also determined that this cleavage of Pseudomonas toxin by furin takes place at the authentic activation site to release the 37-kDa active fragment.
我们已经证明,在完整的中国仓鼠卵巢细胞中,绿脓杆菌外毒素A的天然酶原形式可被弗林蛋白酶在其特定激活位点切割,或者在体外,通过这些细胞分离的膜组分中的弗林蛋白酶进行切割。我们已经将细胞膜组分中弗林蛋白酶的活性与纯化的重组人弗林蛋白酶的活性进行了比较。我们已经证实,RPE.40,一种对绿脓杆菌毒素具有抗性的突变细胞株,在弗林基因中发生了突变,并且我们已经证明这些细胞在毒素切割方面存在缺陷。我们还确定,弗林蛋白酶对绿脓杆菌毒素的这种切割发生在真实的激活位点,以释放37 kDa的活性片段。