Yoon Hyesook, Blaber Sachiko I, Debela Mekdes, Goettig Peter, Scarisbrick Isobel A, Blaber Michael
Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL 32306-4300, USA.
Biol Chem. 2009 Apr;390(4):373-7. doi: 10.1515/BC.2009.026.
We previously reported the activation profiles of the human kallikrein-related peptidases (KLKs) as determined from a KLK pro-peptide fusion-protein system. That report described the activity profiles of 12 of the 15 mature KLKs versus the 15 different pro-KLK sequences. The missing profiles in the prior report, involving KLK9, 10, and 15, are now described. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, mass spectrometry, and N-terminal sequence analyses show that KLK9 and 10 exhibit low hydrolytic activities towards all of the 15 pro-KLK sequences, while KLK15 exhibits significant activity towards both Arg- and Lys-containing KLK pro-sequences. The ability of KLK15 to activate pro-KLK8, 12, and 14 is confirmed using recombinant pro-KLK proteins, and shown to be significant for activation of pro-KLK8 and 14, but not 12. These additional data for KLK9, 10, and 15 now permit a completed KLK activome profile, using a KLK pro-peptide fusion-protein system, to be described. The results suggest that KLK15, once activated, can potentially feed back into additional pro-KLK activation pathways. Conversely, KLK9 and 10, once activated, are unlikely to participate in further pro-KLK activation pathways, although similar to KLK1 they may activate other bioactive peptides.
我们之前报道了通过激肽释放酶原相关肽酶（KLK）前肽融合蛋白系统测定的人KLK的激活谱。该报告描述了15种成熟KLK中的12种相对于15种不同的前激肽释放酶原序列的活性谱。现在描述了先前报告中缺失的涉及KLK9、10和15的谱。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、质谱分析和N端序列分析表明，KLK9和10对所有15种前激肽释放酶原序列均表现出低水解活性，而KLK15对含精氨酸和赖氨酸的激肽释放酶原前序列均表现出显著活性。使用重组前激肽释放酶原蛋白证实了KLK15激活前激肽释放酶原8、12和14的能力，并且表明该能力对激活前激肽释放酶原8和14具有重要意义，但对激活前激肽释放酶原12没有意义。现在，利用KLK前肽融合蛋白系统，这些关于KLK9、10和15的额外数据使得完整的KLK激活组谱得以描述。结果表明，KLK15一旦被激活，可能会反馈到其他前激肽释放酶原激活途径中。相反，KLK9和10一旦被激活，不太可能参与进一步的前激肽释放酶原激活途径，尽管它们与KLK1类似，可能会激活其他生物活性肽。
