Mason R W, Bartholomew L T, Hardwick B S
Department of Biochemistry, Strangeways Research Laboratory, Cambridge, U.K.
Biochem J. 1989 Nov 1;263(3):945-9. doi: 10.1042/bj2630945.
The ability of benzyloxycarbonyl-(125I)Tyr-Ala-CHN2 to label cysteine proteinases in a variety of human tissues was investigated. The inhibitor bound only to cathepsin B in tissues homogenized at pH 5.0. When liver was autolysed at pH 4.0 for up to 4 h, the inhibitor also bound to a protein of Mr 25,000. This was identified immunologically and chromatographically as cathepsin L. Both cathepsins B and L were found primarily in kidney, liver and spleen. In spleen, an additional protein of Mr 25,000 was also labelled. This protein could not be precipitated by antibodies to any of cathepsins B, H and L. This protein has tentatively been identified as human cathepsin S by its tissue distribution, chromatographic properties and molecular size. This work clearly shows that peptidyldiazomethanes are specific probes for cysteine proteinases, and that benzyloxycarbonyl-(125I)Tyr-Ala-CHN2 binds to three such enzymes in human tissues.
研究了苄氧羰基 -（125I）酪氨酸 - 丙氨酸 - 重氮甲烷标记多种人体组织中半胱氨酸蛋白酶的能力。该抑制剂仅在pH 5.0匀浆的组织中与组织蛋白酶B结合。当肝脏在pH 4.0下自溶长达4小时时，该抑制剂还与一种分子量为25,000的蛋白质结合。经免疫和色谱鉴定，该蛋白质为组织蛋白酶L。组织蛋白酶B和L主要存在于肾脏、肝脏和脾脏中。在脾脏中，还标记了一种分子量为25,000的额外蛋白质。该蛋白质不能被针对组织蛋白酶B、H和L的任何一种抗体沉淀出来。根据其组织分布、色谱性质和分子大小，该蛋白质初步被鉴定为人组织蛋白酶S。这项工作清楚地表明，肽基重氮甲烷是半胱氨酸蛋白酶的特异性探针，并且苄氧羰基 -（125I）酪氨酸 - 丙氨酸 - 重氮甲烷在人体组织中可与三种此类酶结合。
Zotero 插件
