Mechanism of Soluble Corin activation by PCSK6

doi:10.13523/j.cb.20160703

China Biotechnology

2016, Vol. 36

Issue (7): 15-20 DOI: 10.13523/j.cb.20160703

Mechanism of Soluble Corin activation by PCSK6

LI Feng^1,2, ZHOU Tian-tian³, LIU Meng³, DONG Ning-zheng^1,2,3

1 The First Affiliated Hospital, Soochow University, Suzhou 215006, China;
2 Thrombosis and Hemostasis Key Laboratory of the Ministry of Health, Jiangsu Institute of Hematology, Suzhou 215006, China;
3 The Cyrus Tang Hematology Center, Soochow University, Suzhou 215123, China

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Abstract

Purpose:To explore whether soluble corin can be activated by PCSK6 (the proprotein convertase subtilisin/kextin type 6) and exhibited biologic activity. Methods:To construct soluble corin expression vector, corin cDNA fragment (463~3219bp) was amplified using pcDNACorin as a template, and cloned into the eukaryotic expression vector pSec/FRT/V5-His-TOPO to yield the plasmid pSECsolCorin. The constructor was verified by restriction endonuclease digestion and DNA sequencing. Then soluble corin, PCSK6 and pro-ANP expression vectors were transfected into HEK293 cells, collected cell lysate and conditioned medium. Western blot were used to determine protein expression. Then soluble corin medium was incubated with increasing amounts of PCSK6 for 1h, activated corin and PCSK6 were detected by Western blot. Then also incubated activated soluble corin with pro-ANP to exam corin activities. Results:Construct soluble corin expression vector, and soluble corin can be expressed in HEK293 cells. When soluble corin was treated with condition medium contain increasing amount PCSK6, activated corin protease fragments (corin-p) can be detected and increased in dose-dependently manner. And activated soluble corin can process pro-ANP to ANP in dose-dependently manner. Conclusions:Soluble corin can be activated by PCSK6, then exhibiting the capacity of processing pro-ANP. Soluble corin may become a new therapeutic strategy of heart failure.

Key words： Soluble corin Heart failure Pro-ANP PCSK6

Received: 01 February 2016 Published: 25 July 2016

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Cite this article:

LI Feng, ZHOU Tian-tian, LIU Meng, DONG Ning-zheng. Mechanism of Soluble Corin activation by PCSK6. China Biotechnology, 2016, 36(7): 15-20.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20160703 OR https://manu60.magtech.com.cn/biotech/Y2016/V36/I7/15

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