基质金属蛋白酶 1 和金属蛋白酶组织抑制剂与盘鲍胶原代谢的关系,Journal of Agricultural and Food Chemistry

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基质金属蛋白酶 1 和金属蛋白酶组织抑制剂与盘鲍胶原代谢的关系
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2022-11-18 , DOI: 10.1021/acs.jafc.2c05931
Yu-Lei Chen _{1,

2} , Ming-Hui Zhang ₁ , Le-Le Su ₁ , Le-Chang Sun _{1,

2} , Xu-Jian Qiu ₁ , Duanquan Lin _{1,

2} , Ling-Jing Zhang _{1,

2} , Tengchuan Jin ₃ , Min-Jie Cao _{1,

2}

Affiliation

为了响应物理、化学和/或生物刺激，鲍鱼会发生相当大的组织自降解，导致严重的收获后质量损失。在此过程中，细胞外基质 (ECM) 被内源性蛋白酶大大降解。ECM 的主要成分是胶原蛋白，主要是 I 型胶原蛋白。虽然可以特异性降解胶原蛋白的基质金属蛋白酶（MMPs）的活性受到MPs组织抑制剂（TIMPs）的精确调节，表明MMPs和TIMPs在组织自降解的调节中起着至关重要的作用，但很少有研究报道MMP 和 TIMP 之间的相互作用。在这项研究中，我们揭示了胶原酶参与Haliotis discus hannai的死后组织自我降解通过降解 I 型胶原蛋白。利用原核表达系统表达了具有高纯度和酶活性的重组鲍鱼MMP-1催化结构域（rMMP1c）。rMMP1c 的最适温度和 pH 值分别为 37 °C 和 7.0。rMMP1c 的热变性温度为 67.0 ± 0.9 °C。乙二胺四乙酸（EDTA）和1,10-菲咯啉可完全抑制rMMP1c活性，而Ba ²⁺、Ca ²⁺和Mg ²⁺可以显着提升它。TIMP 也使用 HEK 293F 细胞表达。重组 TIMP (rTIMP) 对 rMMP1c 显示出良好的抑制活性。抑制动力学分析表明 rTIMP 是 rMMP1c 的竞争性抑制剂。生物层干涉测量显示 rTIMP 可以有效地与 rMMP1c 结合，平衡解离常数值为 263 nM。rMMP1c依次有效降解I型胶原γ-β-α链，rTIMP可显着抑制rMMP1c降解活性。这些结果为研究MMP与TIMP相互作用提供了理论基础，阐明了鲍鱼组织自降解的可能机制。

"点击查看英文标题和摘要"

Relationships of Matrix Metalloproteinase 1 and a Tissue Inhibitor of Metalloproteinase to Collagen Metabolism in Haliotis discus hannai

In response to physical, chemical, and/or biological stimuli, considerable tissue self-degradation occurs in abalone, causing severe post-harvest quality loss. During this process, the extracellular matrix (ECM) is greatly degraded by endogenous proteases. The main component of the ECM is collagen, primarily type I collagen. Although the activity of matrix metalloproteinases (MMPs), which can specifically degrade collagen, is precisely regulated by tissue inhibitors of MPs (TIMPs), indicating that MMPs and TIMPs play crucial roles in the regulation of tissue self-degradation, few studies have reported the interaction between MMPs and TIMPs. In this study, we reveal collagenases to participate in postmortem tissue self-degradation of Haliotis discus hannai by degrading type I collagen. The recombinant MMP-1 catalytic domain (rMMP1c) of abalone with high purity and enzyme activity is expressed using a prokaryotic expression system. The optimum temperature and pH for rMMP1c are 37 °C and 7.0, respectively. The thermal denaturation temperature of rMMP1c is 67.0 ± 0.9 °C. Ethylenediamine tetraacetic acid (EDTA) and 1,10-phenanthroline can completely inhibit rMMP1c activity, while Ba²⁺, Ca²⁺, and Mg²⁺ can significantly elevate it. TIMP is also expressed using HEK 293F cells. Recombinant TIMP (rTIMP) shows good inhibitory activity toward rMMP1c. Inhibition kinetics analyses reveal rTIMP to be a competitive inhibitor of rMMP1c. Biolayer interferometry reveals that rTIMP can effectively bind with rMMP1c, with an equilibrium dissociation constant value of 263 nM. rMMP1c effectively degrades type I collagen γ–β–α chains in turn, and rTIMP can significantly inhibit rMMP1c degradation activity. These results provide a theoretical basis for the study of MMP and TIMP interaction and elucidate the possible mechanism for abalone tissue self-degradation.

更新日期：2022-11-18

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