Cleavage of desmin by cysteine proteases: Calpains and cathepsin B
Cleavage of desmin by cysteine proteases: Calpains and cathepsin B
复制标题
DOI：
10.1016/j.meatsci.2004.03.019
复制
发表时间：
2004-11-01
期刊：
影响因子：
7.1
通讯作者：
Purslow, PP
中科院分区：
文献类型：
--
作者：
Baron, CP;Jacobsen, S;Purslow, PP
从猪背最长肌中纯化中间丝蛋白结蛋白，并与P-钙蛋白酶、m-钙蛋白酶或组织蛋白酶B孵育。随后使用SDS-PAGE和Western印迹法对结蛋白进行蛋白水解。在将结蛋白与蛋白酶孵育后，通过不同蛋白水解片段的N-末端测序鉴定结蛋白分子上的切割位点。与m-钙蛋白酶或mu-钙蛋白酶孵育的结蛋白主要在头部和尾部区域中裂解，即使在长时间孵育后也留下相对完整的杆结构域。与组织蛋白酶B孵育产生该二肽基肽酶特征性的连续C末端降解模式。底物的一级结构没有被发现是必不可少的半胱氨酸肽酶的蛋白水解活性的调节研究。然而，获得的降解模式意味着，钙蛋白酶参与降解结蛋白死后早期，靶向非螺旋区的结蛋白分子，并导致解聚和初始解体的中间丝结构的肌细胞。(C)2004爱思唯尔有限公司保留所有权利。
The intermediate filament protein, desmin, was purified from pork longissimus dorsi and incubated with either P-calpain, m-calpain or cathepsin B. Proteolysis of desmin was followed using SDS-PAGE and Western blotting. After incubation of desmin with the proteases, cleavage sites on the desmin molecule were identified by N-terminal sequencing of the different proteolytic fragments. Desmin incubated with either m-calpain or mu-calpain was primarily cleaved in the head and tail region leaving the rod domain relatively intact even after prolonged incubation. Incubation with cathepsin B produces a sequential C-terminal degradation pattern characteristic of this dipeptylpeptidase. The substrate primary structure was not found to be essential for regulation of the proteolytic activity of the cysteine peptidases studied. However, the degradation patterns obtained imply that calpains are involved in degradation of desmin early post-mortem, targeting the non-helical region of the desmin molecule and resulting in depolymerisation and initial disorganisation of the intermediate filament structures of the muscle cell. (C) 2004 Elsevier Ltd. All rights reserved.