Identification and enzymatic characterization of two diverging murine counterparts of human interstitial collagenase (MMP-1) expressed at sites of embryo implantation
Identification and enzymatic characterization of two diverging murine counterparts of human interstitial collagenase (MMP-1) expressed at sites of embryo implantation
复制标题
DOI：
10.1074/jbc.m007674200
复制
发表时间：
2001-03-30
影响因子：
4.8
通讯作者：
López-Otín, C
中科院分区：
文献类型：
--
作者：
Balbín, M;Fueyo, A;López-Otín, C
小鼠组织中纤维状胶原的重塑已被广泛归因于胶原酶-3（基质金属蛋白酶-13（MMP-13））的活性，胶原酶-3是在该物种中鉴定的主要胶原酶。这一建议主要是基于反复无效的尝试来检测鼠组织中间质胶原酶（MMP-1）的存在，MMP-1是许多物种（包括人类）中的主要胶原酶。在这项工作中，我们已经进行了广泛的筛选小鼠基因组和cDNA文库作为探针的全长cDNA的人MMP-1，我们报告的MMP基因家族的两个新成员，这是包含在集群的MMP基因位于小鼠9号染色体的鉴定。分离的cDNA含有464和463个氨基酸的开放阅读框，同源性为82%，显示了MMPs原型的所有结构特征。序列相似性比较发现，与人间质胶原酶（MMP-1）的同源性最高，新蛋白暂命名为Mcol-A和Mcol-B（鼠胶原酶样e和B），重组蛋白的酶活性分析显示两者都是催化自活化的，但只有Mcol-A能够降解合成肽和I型和II型纤维状胶原。Mcol-A和Mcol-B基因均位于小鼠9号染色体的A1-A2区域，Mcol-A占据与人MMP-1基因座在11 q22处同线的位置。这些新的MMPs在小鼠组织中的表达分析显示，它们主要存在于小鼠胚胎发生过程中，特别是在小鼠滋养层巨细胞中。根据其结构和功能特点，我们建议，至少有一个新的MMP家族成员，Mcol-A，可能发挥作用，在小鼠组织间质胶原酶，并可能代表一个真正的直系同源的人MMP-1。
Remodeling of fibrillar collagen in mouse tissues has been widely attributed to the activity of collagenase-3 (matrix metalloproteinase-13 (MMP-13)), the main collagenase identified in this species. This proposal has been largely based on the repeatedly unproductive attempts to detect the presence in murine tissues of interstitial collagenase (MMP-1), a major collagenase in many species, including humans. In this work, we have performed an extensive screening of murine genomic and cDNA libraries using as probe the full-length cDNA for human MMP-1, We report the identification of two novel members of the MMP gene family which are contained within the cluster of MMP genes located at murine chromosome 9. The isolated cDNAs contain open reading frames of 464 and 463 amino acids and are 82% identical, displaying all structural features characteristic of archetypal MMPs, Comparison for sequence similarities revealed that the highest percentage of identities was found with human interstitial collagenase (MMP-1), The new proteins were tentatively called Mcol-A and Mcol-B (Murine collagenase-like e and B), Analysis of the enzymatic activity of the recombinant proteins revealed that both are catalytically autoactivable but only Mcol-A is able to degrade synthetic peptides and type I and II fibrillar collagen. Both Mcol-A and Mcol-B genes are located in the A1-A2 region of mouse chromosome 9, Mcol-A occupying a position syntenic to the human MMP-1 locus at 11q22. Analysis of the expression of these novel MMPs in murine tissues revealed their predominant presence during mouse embryogenesis, particularly in mouse trophoblast giant cells. According to their structural and functional characteristics, we propose that at least one of these novel members of the MMP family, Mcol-A, may play roles as interstitial collagenase in murine tissues and could represent a true orthologue of human MMP-1.