Neutral proteinases of human spleen. Purification and criteria for homogeneity of elastase and cathepsin G.
Neutral proteinases of human spleen. Purification and criteria for homogeneity of elastase and cathepsin G.
复制标题
人脾脏的中性蛋白酶。
作者：
P. Starkey;A. Barrett
1.人脾被发现含有蛋白酶活性对偶氮酪蛋白在中性和碱性pH值。2.高离子强度和某些洗涤剂对活性有促进作用。3.用含0.1%Brij 35和0.1%EDTA三钠的1. 0 M-NaCl溶液提取蛋白酶效果最佳。4.在纯化的初始阶段，在不存在盐的情况下，蛋白酶被有效地吸附到不溶性物质上。5.用DEAE-纤维素层析法分离出两种不同的蛋白酶，一种是弹性蛋白酶，另一种是胰凝乳蛋白酶样酶，称为组织蛋白酶G。6.两种酶通过进一步的柱层析高度纯化。7.通过凝胶色谱和十二烷基硫酸钠凝胶电泳估计酶的分子量。8.通过等电聚焦和凝胶电泳表明，这两种酶都是以多种形式存在的阳离子蛋白。
1. Human spleen was found to contain proteinases active against azo-casein at neutral and alkaline pH values. 2. The activity was stimulated by high ionic strength and some detergents. 3. Optimal extraction of the proteinases from the tissue was achieved with 1.0M-NaCl containing 0.1% Brij 35 and 0.1% trisodium EDTA. 4. The proteinases were efficiently adsorbed to insoluble material in the absence of salt in the initial stages of purification. 5. Two distinct proteinases were separated by chromatography on DEAE-cellulose, an elastase and a chymotrypsin-like enzyme designated cathepsin G. 6. Both enzymes were highly purified by further column chromatography. 7. The molecular weights of the enzymes were estimated by gel chromatography and sodium dodecyl sulphate-gel electrophoresis. 8. It was shown by isoelectric focusing and gel electrophoresis that both enzymes are cationic proteins that occur in multiple forms.