Purification and characterization of Protein C activator from Agkistrodon acutus Venom

Abstract

Protein C (PC) plays an important role in the balance of coagulation and anticoagulation. Thus, the detection of PC activity is diagnostically significant for patients with cardiovascular diseases. Presently, the key methods to detect PC activity are the chromogenic assay and activated partial thromboplastin time (APTT) test. PROTAC used in the chromogenic assay is isolated from Agkistrodon contortrix venom as protein C activator (PCA). However, the use of the chromogenic assay is limited because of the high price of PROTAC. In this study, PCA was successfully purified from Agkistrodon acutus venom (AAV) by ion-exchange and gel chromatography. PCA from AAV has a relative molecular mass of 24 kD, calculated from the measurement of 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The components of PCA were identified by MALDI-TOF/TOF-MS and mascot searches revealed that the coverage rate between PCA and zinc metalloproteinase AaPA from AAV was 21%. The chromogenic assay and APTT test were used to measure the enzymatic activity of PCA, and the results showed that PCA from AAV could specifically activate PC. In summary, the chromogenic assay described herein is highly sensitive and easy to perform.

Keywords:

• Agkistrodon acutus venom
• Protein C
• Protein C activator

Disclosure statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

We acknowledge Drs. Zheng Shu-Guo, Yan Liang, and Xu Lei for writing assistance as well as Ms. Jiang Meng for providing language help. We thank the Shanghai Institute of Biological Sciences, Chinese Academy of Sciences, for performing the protein mass spectrometry experiment. We thank TopEdit (www.topeditsci.com) for its linguistic assistance during the preparation of this manuscript.

Correction Statement

This article has been republished with minor changes. These changes do not impact the academic content of the article.

Additional information

Funding

This work was supported by grants from the provincial natural science fund of Anhui (KJ2011A266), the key natural science cultivation fund of Wannan Medical College (WK2016Z02) and the key natural science fund of Wannan Medical College (WK2019Z09).