Activation profiles of human kallikrein-related peptidases by proteases of the thrombostasis axis.
Activation profiles of human kallikrein-related peptidases by proteases of the thrombostasis axis.
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血栓形成轴蛋白酶对人激肽释放酶相关肽酶的激活谱。
DOI：
10.1110/ps.036715.108
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Blaber,Michael
中科院分区：
文献类型：
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作者：
Yoon,Hyesook;Blaber,SachikoI;Evans,DMichael;Trim,Julie;Juliano,MariaAparecida;Scarisbrick,IsobelA;Blaber,Michael
人激肽释放酶相关多肽酶(KLKs)由15个成员组成(KLK1-15)，是最大的丝氨酸蛋白酶家族。KLK被用作或建议用作癌症和神经退行性疾病的重要临床生物标志物和治疗靶点。所有的KLK似乎都是以非活性前体(PRO-KLK)的形式分泌出来的，通过其N-末端前肽的特异性蛋白水解性释放在细胞外被激活。这一过程是调节KLK功能的关键步骤。最近的许多工作都致力于阐明KLK家族成员之间激活级联的可能性，目前在皮肤脱皮和精液液化中描述了与生理相关的KLK调节级联。尽管对KLK调控的了解越来越多，但关于KLK与其他调节蛋白水解酶功能交叉的可能性的细节基本上是未知的。为了阐明这种相互作用的可能性，我们使用先前描述的前KLK融合蛋白系统表征了与血栓形成相关的蛋白水解酶对KLK家族前肽序列的降解能力。随后，使用完整的重组原KLK蛋白进行蛋白水解性分析，对部分阳性水解物结果进行量化。纤溶酶和uPA都能激活原KLK6和14，其中纤溶酶是目前发现的最好的原KLK6激活剂。PRO-KLK11和PRO-KLK12可以被广泛的凝血酶激活，凝血酶对PRO-KLK12具有高度的选择性。结果表明，血栓形成蛋白家族的蛋白可以有效地激活特定的原-KLK，表明这两个主要的蛋白水解酶家族之间可能存在重要的调节相互作用。
The human kallikrein‐related peptidases (KLKs) comprise 15 members (KLK1–15) and are the single largest family of serine proteases. The KLKs are utilized, or proposed, as clinically important biomarkers and therapeutic targets of interest in cancer and neurodegenerative disease. All KLKs appear to be secreted as inactive pro‐forms (pro‐KLKs) that are activated extracellularly by specific proteolytic release of their N‐terminal pro‐peptide. This processing is a key step in the regulation of KLK function. Much recent work has been devoted to elucidating the potential for activation cascades between members of the KLK family, with physiologically relevant KLK regulatory cascades now described in skin desquamation and semen liquefaction. Despite this expanding knowledge of KLK regulation, details regarding the potential for functional intersection of KLKs with other regulatory proteases are essentially unknown. To elucidate such interaction potential, we have characterized the ability of proteases associated with thrombostasis to hydrolyze the pro‐peptide sequences of the KLK family using a previously described pro‐KLK fusion protein system. A subset of positive hydrolysis results were subsequently quantified with proteolytic assays using intact recombinant pro‐KLK proteins. Pro‐KLK6 and 14 can be activated by both plasmin and uPA, with plasmin being the best activator of pro‐KLK6 identified to date. Pro‐KLK11 and 12 can be activated by a broad‐spectrum of thrombostasis proteases, with thrombin exhibiting a high degree of selectivity for pro‐KLK12. The results show that proteases of the thrombostasis family can efficiently activate specific pro‐KLKs, demonstrating the potential for important regulatory interactions between these two major protease families.