牛胰脏纯化组织蛋白酶L对体外肌内结缔组织的降解作用
The Degradation of Intramuscular Connective Tissue In Vitro with Purified Cathepsin L from Bovine Pancreas.
作者信息
Peng Yingbo, He Wanhong, Teng Shuang, Jamali Muneer Ahmed
机构信息
College of Engineering, Nanjing Agricultural University, Nanjing 210095, China.
College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
出版信息
Foods. 2023 Sep 21;12(18):3517. doi: 10.3390/foods12183517.
To investigate the possible degradation of the intramuscular connective tissue (IMCT) with cathepsin L, isolated IMCTs were incubated with purified cathepsin L in vitro. Here, we prepared purified cathepsin L from bovine pancreas by using DEAE Sephacel, Sephacryl S-100 HR, SP Sepharose FF, and con A-Sepharose affinity chromatography in sequence. An SDS-PAGE analysis of CNBr-digested peptides showed that the degradation of collagen in IMCT could take place on terminal non-helical peptides rather than the triple helix region. Decorin (DCN) was clearly degraded at a pH of 5.0. The T and T of intramuscular connective tissue decreased to 41.41 °C and 43.79 °C, respectively. In the cathepsin L treatment of pH 5.0, the decreases in the T and T of IMCT were more sensitive than they were at pH 5.5~6.5.
为研究组织蛋白酶L对肌内结缔组织（IMCT）的潜在降解作用，将分离出的IMCTs与纯化的组织蛋白酶L在体外进行孵育。在此，我们依次使用DEAE Sephacel、Sephacryl S - 100 HR、SP Sepharose FF和伴刀豆球蛋白A - Sepharose亲和层析从牛胰腺中制备纯化的组织蛋白酶L。对溴化氰消化肽段的SDS - PAGE分析表明，IMCT中胶原蛋白的降解可能发生在末端非螺旋肽段而非三螺旋区域。在pH为5.0时，核心蛋白聚糖（DCN）明显降解。肌内结缔组织的热变性温度T₀和热焓变ΔH分别降至41.41℃和43.79℃。在pH 5.0的组织蛋白酶L处理中，IMCT的T₀和ΔH的降低比在pH 5.5至6.5时更敏感。
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