The purpose of this study was to analyze spatio-temporal dynamics of localization of protease-sensitive sites Arg-X in non-histone and histone blocks of heteropolymer suprastructures （nucleoplasm, chromatin, nuclear ...The purpose of this study was to analyze spatio-temporal dynamics of localization of protease-sensitive sites Arg-X in non-histone and histone blocks of heteropolymer suprastructures （nucleoplasm, chromatin, nuclear matrix） as possible zones affecting the conformational rearrangements of the total interphase chromatin at the induction of increasing morphogenesis of mature embryos-germs of spring and transformed from its winter wheat. Germinated embryos-germs were detached from endosperm after 24 hours from the start of soaking. Cell nuclei have been allocated from embryos-germs and cleared, and then from their heteropolymer suprastructures （nucleoplasm, chromatin loosely bound with nuclear matrix and chromatin tightly bound with nuclear matrix, and nuclear matrix） were extracted by increasing ionic strength of solution. From isolated nuclear suprastructures, non-histone proteins were separated from histones using ion exchange chromatography. Trypsin-like complexes from non-histone proteins and histone blocks were isolated using the affinity chromatography. The Arg-X （tryptase） activity was assessed by cleavage of Arg-X bonds in the arginine-enriched protein protamine. Hypersensitivity to the Arg-X proteolysis in trypsin-like complexes detected at the level suprastructures of chromatin tightly bound with the nuclear matrix was shown. The most active changes of the nuclear proteome have occurred at the level of the non-histone proteins and the core histones （H2A ＋ H2B） （H3 ＋ H4） of induced to growth embryos-seedlings of winter wheat （compared to the initial spring form of wheat）. Perhaps hypersensitivity to the Arg-X activity of the trypsin-like complexes in the non-histone proteins and the core blocks of chromatin tightly bound with nuclear matrix have been entrenched during the transforming of the winter wheat from the initial spring wheat.展开更多
小麦纹枯病菌(Rhizoctonia cerealisVanderHoeven)在以小麦细胞壁为唯一碳氮源的液体培养基中可产生多种蛋白酶,培养液经硫酸铵沉淀,SP-Sepharose Fast F low离子交换层析,Pheny l(high-sub)-Sepharose Fast F low疏水层析,SephadexG-7...小麦纹枯病菌(Rhizoctonia cerealisVanderHoeven)在以小麦细胞壁为唯一碳氮源的液体培养基中可产生多种蛋白酶,培养液经硫酸铵沉淀,SP-Sepharose Fast F low离子交换层析,Pheny l(high-sub)-Sepharose Fast F low疏水层析,SephadexG-75分子筛层析,分离纯化到分子量约40 kD的一种蛋白酶。该酶能水解胰蛋白酶专一性底物Benz-Phe-Val-A rg-NA,对凝乳弹性蛋白酶底物Suc-A la-A la-Pro-Phe-NA和枯草杆菌蛋白酶底物CBZ-G ly-G ly-Leu-NA无活性,能被胰蛋白酶抑制剂aprotinin、leupeptin和soybean trypsin inhib itor强烈抑制,丝氨酸蛋白酶抑制剂PM SF能部分抑制酶的活性,表明该蛋白酶是丝氨酸蛋白酶中的一种类胰蛋白酶。展开更多
目的:观察灭蚊真菌贵阳腐霉产生类胰蛋白酶(Pr2)所需的诱导条件。方法:通过单因素诱导物、温度、pH值等比较试验摸索其产生Pr2的适宜条件。结果:贵阳腐霉产Pr2的适宜条件是在100 m l基本盐培养基中,以1%蝉蜕为唯一碳氮源,在26℃,初始pH...目的:观察灭蚊真菌贵阳腐霉产生类胰蛋白酶(Pr2)所需的诱导条件。方法:通过单因素诱导物、温度、pH值等比较试验摸索其产生Pr2的适宜条件。结果:贵阳腐霉产Pr2的适宜条件是在100 m l基本盐培养基中,以1%蝉蜕为唯一碳氮源,在26℃,初始pH值8.0,1%的菌丝接种量。结论:本研究初步确定的贵阳腐霉的Pr2产酶条件,为该酶的进一步研究提供了有价值的参考。展开更多
文摘The purpose of this study was to analyze spatio-temporal dynamics of localization of protease-sensitive sites Arg-X in non-histone and histone blocks of heteropolymer suprastructures （nucleoplasm, chromatin, nuclear matrix） as possible zones affecting the conformational rearrangements of the total interphase chromatin at the induction of increasing morphogenesis of mature embryos-germs of spring and transformed from its winter wheat. Germinated embryos-germs were detached from endosperm after 24 hours from the start of soaking. Cell nuclei have been allocated from embryos-germs and cleared, and then from their heteropolymer suprastructures （nucleoplasm, chromatin loosely bound with nuclear matrix and chromatin tightly bound with nuclear matrix, and nuclear matrix） were extracted by increasing ionic strength of solution. From isolated nuclear suprastructures, non-histone proteins were separated from histones using ion exchange chromatography. Trypsin-like complexes from non-histone proteins and histone blocks were isolated using the affinity chromatography. The Arg-X （tryptase） activity was assessed by cleavage of Arg-X bonds in the arginine-enriched protein protamine. Hypersensitivity to the Arg-X proteolysis in trypsin-like complexes detected at the level suprastructures of chromatin tightly bound with the nuclear matrix was shown. The most active changes of the nuclear proteome have occurred at the level of the non-histone proteins and the core histones （H2A ＋ H2B） （H3 ＋ H4） of induced to growth embryos-seedlings of winter wheat （compared to the initial spring form of wheat）. Perhaps hypersensitivity to the Arg-X activity of the trypsin-like complexes in the non-histone proteins and the core blocks of chromatin tightly bound with nuclear matrix have been entrenched during the transforming of the winter wheat from the initial spring wheat.
文摘小麦纹枯病菌(Rhizoctonia cerealisVanderHoeven)在以小麦细胞壁为唯一碳氮源的液体培养基中可产生多种蛋白酶,培养液经硫酸铵沉淀,SP-Sepharose Fast F low离子交换层析,Pheny l(high-sub)-Sepharose Fast F low疏水层析,SephadexG-75分子筛层析,分离纯化到分子量约40 kD的一种蛋白酶。该酶能水解胰蛋白酶专一性底物Benz-Phe-Val-A rg-NA,对凝乳弹性蛋白酶底物Suc-A la-A la-Pro-Phe-NA和枯草杆菌蛋白酶底物CBZ-G ly-G ly-Leu-NA无活性,能被胰蛋白酶抑制剂aprotinin、leupeptin和soybean trypsin inhib itor强烈抑制,丝氨酸蛋白酶抑制剂PM SF能部分抑制酶的活性,表明该蛋白酶是丝氨酸蛋白酶中的一种类胰蛋白酶。
文摘目的:观察灭蚊真菌贵阳腐霉产生类胰蛋白酶(Pr2)所需的诱导条件。方法:通过单因素诱导物、温度、pH值等比较试验摸索其产生Pr2的适宜条件。结果:贵阳腐霉产Pr2的适宜条件是在100 m l基本盐培养基中,以1%蝉蜕为唯一碳氮源,在26℃,初始pH值8.0,1%的菌丝接种量。结论:本研究初步确定的贵阳腐霉的Pr2产酶条件,为该酶的进一步研究提供了有价值的参考。
