Carolina Proaño-Bolaños , Renjie Li , Mei Zhou , Lei Wang , Xinping Xi , Elicio E. Tapia , Luis A. Coloma , Tianbao Chen , Chris Shaw
{"title":"从金叶蛙皮肤分泌物中提取新的卡扎尔型蛋白酶抑制剂","authors":"Carolina Proaño-Bolaños , Renjie Li , Mei Zhou , Lei Wang , Xinping Xi , Elicio E. Tapia , Luis A. Coloma , Tianbao Chen , Chris Shaw","doi":"10.1016/j.euprot.2017.02.001","DOIUrl":null,"url":null,"abstract":"<div><p>Peptidase inhibitors have an important role controlling a variety of biological processes. Here, we employed a peptidomic approach including molecular cloning, tandem mass spectrometry and enzymatic assays to reveal 7 Kazal-type proteinase inhibitors (CCKPs) (18 variants) in the skin secretion of the unexplored frog, <em>Cruziohyla calcarifer</em>. All 18 proteins shared the Kazal pattern C-X<sub>(7)</sub>-C-X<sub>(6,7)</sub>-C-X<sub>(6,7)</sub>-Y-X<sub>(3)</sub>-C-X<sub>(2)</sub>-C-X<sub>(15-21)</sub>-C and 3 disulphide bridges. Based on structural comparative analysis, we deemed trypsin and chymotrypsin inhibitory activity in CCKP-1, 4 and CCKP 2, 5, 7, respectively. These peptidase inhibitors presumably play a role to control the balance between other functional peptides produced in the amphibian skin secretions.</p></div>","PeriodicalId":38260,"journal":{"name":"EuPA Open Proteomics","volume":"15 ","pages":"Pages 1-13"},"PeriodicalIF":0.0000,"publicationDate":"2017-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.euprot.2017.02.001","citationCount":"8","resultStr":"{\"title\":\"Novel Kazal-type proteinase inhibitors from the skin secretion of the Splendid leaf frog, Cruziohyla calcarifer\",\"authors\":\"Carolina Proaño-Bolaños , Renjie Li , Mei Zhou , Lei Wang , Xinping Xi , Elicio E. Tapia , Luis A. Coloma , Tianbao Chen , Chris Shaw\",\"doi\":\"10.1016/j.euprot.2017.02.001\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Peptidase inhibitors have an important role controlling a variety of biological processes. Here, we employed a peptidomic approach including molecular cloning, tandem mass spectrometry and enzymatic assays to reveal 7 Kazal-type proteinase inhibitors (CCKPs) (18 variants) in the skin secretion of the unexplored frog, <em>Cruziohyla calcarifer</em>. All 18 proteins shared the Kazal pattern C-X<sub>(7)</sub>-C-X<sub>(6,7)</sub>-C-X<sub>(6,7)</sub>-Y-X<sub>(3)</sub>-C-X<sub>(2)</sub>-C-X<sub>(15-21)</sub>-C and 3 disulphide bridges. Based on structural comparative analysis, we deemed trypsin and chymotrypsin inhibitory activity in CCKP-1, 4 and CCKP 2, 5, 7, respectively. These peptidase inhibitors presumably play a role to control the balance between other functional peptides produced in the amphibian skin secretions.</p></div>\",\"PeriodicalId\":38260,\"journal\":{\"name\":\"EuPA Open Proteomics\",\"volume\":\"15 \",\"pages\":\"Pages 1-13\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2017-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/j.euprot.2017.02.001\",\"citationCount\":\"8\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"EuPA Open Proteomics\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S221296851630037X\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"EuPA Open Proteomics","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S221296851630037X","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 8
摘要
肽酶抑制剂在控制多种生物过程中起着重要作用。在这里,我们采用肽组学方法,包括分子克隆、串联质谱和酶分析,揭示了未被发现的蛙Cruziohyla calcarifer皮肤分泌物中的7种kazal型蛋白酶抑制剂(CCKPs)(18种变体)。所有18个蛋白都具有卡扎尔模式C-X(7)-C- x (6,7)-C- x (6,7)-Y-X(3)-C- x (2)-C- x (15-21)-C和3个二硫桥。基于结构对比分析,我们认为CCKP-1、4和CCKP- 2、5、7分别具有胰蛋白酶和凝乳胰蛋白酶抑制活性。这些肽酶抑制剂可能在控制两栖动物皮肤分泌物中产生的其他功能肽之间的平衡中起作用。
Novel Kazal-type proteinase inhibitors from the skin secretion of the Splendid leaf frog, Cruziohyla calcarifer
Peptidase inhibitors have an important role controlling a variety of biological processes. Here, we employed a peptidomic approach including molecular cloning, tandem mass spectrometry and enzymatic assays to reveal 7 Kazal-type proteinase inhibitors (CCKPs) (18 variants) in the skin secretion of the unexplored frog, Cruziohyla calcarifer. All 18 proteins shared the Kazal pattern C-X(7)-C-X(6,7)-C-X(6,7)-Y-X(3)-C-X(2)-C-X(15-21)-C and 3 disulphide bridges. Based on structural comparative analysis, we deemed trypsin and chymotrypsin inhibitory activity in CCKP-1, 4 and CCKP 2, 5, 7, respectively. These peptidase inhibitors presumably play a role to control the balance between other functional peptides produced in the amphibian skin secretions.