Structural Characterization of Mouse Neutrophil Serine Proteases and Identification of Their Substrate Specificities RELEVANCE TO MOUSE MODELS OF HUMAN INFLAMMATORY DISEASES
Structural Characterization of Mouse Neutrophil Serine Proteases and Identification of Their Substrate Specificities RELEVANCE TO MOUSE MODELS OF HUMAN INFLAMMATORY DISEASES
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DOI：
10.1074/jbc.m109.042903
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发表时间：
2009-12-04
影响因子：
4.8
通讯作者：
Gauthier, Francis
中科院分区：
文献类型：
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作者：
Kalupov, Timofey;Brillard-Bourdet, Michele;Gauthier, Francis
中性粒细胞丝氨酸蛋白酶（NSP）在嗜中性粒细胞相关的肺部炎症和组织破坏性疾病中起着重要作用。为了研究NSP的致病作用，已经开发了模拟急性或慢性损伤的人肺的各种小鼠实验模型。我们和其他人正在使用暴露于香烟烟雾的小鼠作为慢性阻塞性肺疾病加重或不加重的模型。然而，NSP对肺部疾病过程的相对贡献及其潜在机制仍然知之甚少。缺乏纯化的小鼠NSPs及其特异性底物阻碍了这些研究的进展。在这项工作中，我们比较了小鼠和人类的NSPs，并根据其人类同源物的三维结构生成了小鼠NSPs的三维模型。这些模型的分析提供了令人信服的证据，肽底物特异性的人类和小鼠的NSP是不同的，尽管他们保守的裂缝和密切的结构相似性。这些研究使我们能够首次合成用于单个小鼠NSP的新型灵敏荧光共振能量转移底物。我们的研究结果和新发现的底物应该更好地了解NSP在COPD相关的慢性阻塞性肺疾病以及其他COPD相关的炎症性疾病的发病机制中的作用。
It is widely accepted that neutrophil serine proteases (NSPs) play a critical role in neutrophil-associated lung inflammatory and tissue-destructive diseases. To investigate NSP pathogenic role(s), various mouse experimental models have been developed that mimic acutely or chronically injured human lungs. We and others are using mouse exposure to cigarette smoke as a model for chronic obstructive pulmonary disease with or without exacerbation. However, the relative contribution of NSPs to lung disease processes as well as their underlying mechanisms remains still poorly understood. And the lack of purified mouse NSPs and their specific substrates have hampered advances in these studies. In this work, we compared mouse and human NSPs and generated three-dimensional models of murine NSPs based on three-dimensional structures of their human homologs. Analyses of these models provided compelling evidence that peptide substrate specificities of human and mouse NSPs are different despite their conserved cleft and close structural resemblance. These studies allowed us to synthesize for the first time novel sensitive fluorescence resonance energy transfer substrates for individual mouse NSPs. Our findings and the newly identified substrates should better our understanding about the role of NSPs in the pathogenesis of cigarette-associated chronic obstructive pulmonary disease as well as other neutrophils-associated inflammatory diseases.